Antiproliferative quinazolines

ABSTRACT

Quinazoline compounds which demonstrate antiproliferative activity, such as antitumor activity, processes of preparing these compounds, pharmaceutical compositions containing these compounds, and the use of these compounds. These compounds inhibit the growth and proliferation of the cells of higher organisms and microorganisms, such as bacteria, yeasts and fungi. Preferred quinazoline compounds are capable of inhibiting the enzyme thymidylate synthase. Effects derived from the inhibition of the enzyme thymidylate synthase include those discussed above.

FIELD OF THE INVENTION

The present invention relates to certain quinazoline compounds whichdemonstrate antiproliferative activity, such as antitumor activity, toprocesses for preparing these compounds, to pharmaceutical compositionscontaining these compounds, and to the use of these compounds to inhibitthe growth and proliferation of the cells of higher organisms andmicroorganisms, such as bacteria, yeasts and fungi. Preferred compoundsof the present invention are capable of inhibiting the enzymethymidylate synthase. Effects derived from the inhibition of the enzymethymidylate synthase include those discussed above.

BACKGROUND OF THE INVENTION

A large class of antiproliferative agents includes antimetabolitecompounds. A particular subclass of antimetabolites known as antifolatesor antifols are antagonists of the vitamin folic acid. Typically,antifolates closely resemble the structure of folic acid and incorporatethe characteristic p-benzoyl glutamate moiety of folic acid. Theglutamate moiety of folic acid takes on a double negative charge atphysiological pH. Therefore, this compound and its analogues have anactive, energy-driven transport system to cross the cell membrane andexert a metabolic effect. On the other hand, a compound without theglutamate group may passively diffuse into a

A valid target for an antifolate is the enzyme thymidylate synthase.Thymidylate synthase catalyzes the C-methylation of 2'-deoxyuridylate("dUMP") to provide 2'-deoxythymidylate ("dTMP"). This one-carbontransfer reaction is critical to cell division. Thus, a number of folateanalogues have been synthesized and studied for their ability to inhibitthe enzyme thymidylate synthase. A prototypic, specific, tight-bindinginhibitor of thymidylate synthase, 10-propargyl-5,8-dideazafolic acid(T. R. Jones et al., "A Potent Antitumor Quinazoline Inhibitor ofThymidylate Synthetase: Synthesis, Biological Properties and TherapeuticResults in Mice," Eur. J. Cancer 17:11 (1981)), has shown activityagainst ovarian, liver and breast cancer, with, however, troublesomehepatic and renal toxicities (A. H. Calvert et al., "A Phase IEvaluation of the Quinazoline Antifolate Thymidylate Synthase Inhibitor,N10-Propargyl-5,8-Dideazafolic Acid, CB3717," J. Clin. Oncol. 4:1245(1986)). By addressing two properties in this class of molecule(solubility and capability for intracellular polyglutamation), asuperior second generation analogue (ICI D1694) was developed.

Several lipophilic thymidylate synthase inhibitors have been developedrecently. (See, e.g., E. M. Berman et al., "Substituted Quinazolinonesas Anticancer Agents," U.S. Pat. No. 4,857,530; T. R. Jones et al.,"Antiproliferative Cyclic Compounds," Copending U.S. patent applicationSer. No. 07/432,338, which is a continuation application of Ser. No.07/251,765 filed Sep. 30, 1988; M. D. Varney et al., "AntiproliferativeSubstituted Naphthalene Compounds," U.S. patent application Ser. No.07/583,970 filed Sep. 17, 1990; S. H. Reich et al., "AntiproliferativeSubstituted Tricyclic Compounds," U.S. patent application Ser. No.07/587,666 filed Sep. 25, 1990; L. R. Hughes et al., "Anti-tumourAgents," European Patent Application No. 373891, filed Dec. 12, 1989;and T. R. Jones et al., "Antifolate Quinazolines," U.S. patentapplication Ser. No. 07/812,274 filed Dec. 20, 1991).

SUMMARY OF THE INVENTION

The present invention relates to novel quinazoline compounds whichdemonstrate antiproliferative activity, such as antitumor activity.These compounds are effective in inhibiting the growth and proliferationof the cells of higher organisms and of microorganisms, such asbacteria, yeasts and fungi, processes for preparing these compounds,pharmaceutical compositions containing these compounds, and the use ofthese compounds. Preferred quinazoline compounds according to thepresent invention are capable of inhibiting the enzyme thymidylatesynthase. Effects derived from the inhibition of the enzyme thymidylatesynthase include those discussed above.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to quinazoline compounds having theformula I ##STR1## wherein: R¹ represents hydrogen, halogen, alkyl,--OH, --O-alkyl, --O-(aryl or heteroaryl), --S-alkyl, --S-(aryl orheteroaryl), --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO, --NHOH,--NHO-alkyl, --NHNH₂, substituted --NHNH₂, --NHC(═NH)NH₂,--NHC(═NH)alkyl, fluoroalkyl, cycloalkyl, alkenyl, alkynyl, aryl, orheterocycle;

R² and R³, which may be the same or different, represent hydrogen,halogen, alkyl, cycloalkyl, --OH, --O-alkyl, --S-alkyl, --NH₂,--NH-alkyl, --N-(alkyl)₂, --NHCHO, --NO₂, --NHOH, --NHO-alkyl, --NHNH₂,substituted --NHNH₂, --CN, --CO₂ H, --CO₂ -alkyl, --CONH₂, --CONH-alkyl,--CON(alkyl)₂, --CSNH₂, --CSNH-alkyl, --CSN(alkyl)₂, --C(═NH)NH₂,--NHC(═NH)NH₂, --NHC(═NH)alkyl, --SO-alkyl, --SO₂ -alkyl, fluoroalkyl,--O-fluoroalkyl, --S-fluoroalkyl, --NHCO(alkyl), --NHCO(fluoroalkyl),--SO-fluoroalkyl, --SO₂ -fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂NH(alkyl), --SO₂ N (alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle;

Z represents O or S;

R⁴ represents O, S, SO, SO₂, NH, N-alkyl, CH₂, CH-alkyl, CH-(aryl orheteroaryl), CHOH, CHO-alkyl, CHO-(aryl or heteroaryl), C(alkyl)₂,C(aryl or heteroaryl)₂, C(alkyl)(aryl or heteroaryl), CHS-alkyl,CHS-(aryl or heteroaryl), C(OH)(alkyl), C(OH)(aryl or heteroaryl),C(OH)(cycloalkyl), N(OH), N-cycloalkyl, N(aryl or heteroaryl),C(cycloalkyl)₂, C(aryl or heteroaryl)(cycloalkyl), C(alkyl)(alkenyl),C(alkyl)(alkynyl), C(alkenyl)₂, C(alkynyl)₂, C(alkynyl)(aryl orheteroaryl), C(alkynyl)(alkenyl), C(alkenyl)(aryl or heteroaryl),C(cycloalkyl)(alkenyl), C(cycloalkyl)(alkynyl), C(alkyl)(aryl orheteroaryl), CH(cycloalkyl), CH(alkenyl), CH(alkynyl),C(alkyl)(cycloalkyl), C(alkyl)(O-alkyl), C(alkenyl)(O-alkyl),C(alkynyl)(O-alkyl), C(alkyl)(O-cycloalkyl), C(alkenyl)(O-cycloalkyl),C(alkynyl)(O-cycloalkyl), C(aryl or heteroaryl)(O-alkyl), C(aryl orheteroaryl)(O-cycloalkyl), C(alkynyl)(S-alkyl),C(alkynyl)(S-cycloalkyl), C(alkenyl)(S-alkyl), C(alkenyl)(S-cycloalkyl),C(alkyl)(S-alkyl), C(alkyl)(S-cycloalkyl), C(aryl orheteroaryl)(S-alkyl), C(aryl or heteroaryl)(S-cycloalkyl), N(NH₂),N[NH(alkyl)], N[N(alkyl)₂ ], N[NH(cycloalkyl)], N[N(alkyl)(cycloalkyl)],CH(NH₂), CH[NH(alkyl)], CH[NH(cycloalkyl)], CH[N(alkyl)₂ ],CH[N(alkyl)(cycloalkyl)], CH[N(cycloalkyl)₂ ], C(alkyl)(NH₂),C(alkyl)[NH(alkyl)], C(alkyl)[N(cycloalkyl)₂ ],C(alkyl)[NH(cycloalkyl)], C(alkyl)[N(alkyl)₂ ],C(alkyl)[N(alkyl)(cycloalkyl)], C(aryl or heteroaryl)(NH₂), C(aryl orheteroaryl)]NH(alkyl)], C(aryl or heteroaryl)[NH(cycloalkyl)], C(aryl orheteroaryl)[N(alkyl₂)], C(aryl or heteroaryl)[N(cycloalkyl)₂ ], orC(aryl or heteroaryl)[N(alkyl)(cycloalkyl)]; and

R⁵ represents a substituted or unsubstituted aryl or heteroaryl group.

As used herein, the language "capable of inhibiting the enzymethymidylate synthase," or the like, refers to a compound having athymidylate synthase inhibition constant ("TS K_(i) ") of less than orequal to about 10⁻⁴ M. Preferred compounds according to the presentinvention have TS K_(i) values in the range of less than about 10⁻⁵ M,more preferably less than about 10⁻⁶ M, and most preferably less thanabout 10⁻⁷ M.

Thymidylate synthase is merely exemplary of the activity of thequinazoline compounds of the present invention. Indeed, certaincompounds may demonstrate an antifolate activity besides, or even inaddition to, thymidylate synthase inhibition. Further, certain compoundsmay show antiproliferative activity stemming from a completely differentlocus of action than the inhibition of folic metabolic pathways.

Certain quinazoline compounds according to the present invention maypossess one or more assymetric carbon atoms, and therefore may exist inracemic and optically active forms. The present invention thus isintended to encompass the racemic forms of the quinazoline compoundsaccording to the present invention, as well as any optically activeforms thereof, which possess antitumor activity.

As used herein, the language "alkyl" includes both straight and branchedalkyl groups. An analogous convention applies to other generic termssuch as "alkenyl", "alkynyl" and the like. Furthermore, as used herein,the language "alkyl", "alkenyl", "alkynyl" and the like encompasses bothsubstituted and unsubstituted groups.

The language "alkyl" refers to groups having one to eight, preferablyone to six carbon atoms. For example, "alkyl" may refer to methyl,ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl,pentyl, isopentyl tert-pentyl, hexyl, isohexyl, and the like. Suitablesubstituted alkyls include, but are not limited to, fluoromethyl,difluoromethyl, trifluoromethyl, 2-fluoroethyl, 3-fluoropropyl,hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, and the like.

The language "alkenyl" refers to groups having two to eight, preferablytwo to six carbon atoms. For example, "alkenyl" may refer toprop-2-enyl, but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl,hex-5-enyl, 2,3-dimethylbut-2-enyl, and the like. The language"alkynyl," which also refers to groups having two to eight, preferablytwo to six carbons, includes, but is not limited to, prop-2-ynyl,but-2-ynyl, but-3-ynyl, pent-2-ynyl, 3-methylpent-4-ynyl, hex-2-ynyl,hex-5-ynyl, and the like.

The term "cycloalkyl" as used herein refers to groups having three toseven, preferably three to six carbon atoms. Suitable cycloalkylsinclude, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl and the like. The language "heterocycle," whichrefers to groups having one or more heteroatoms, and preferably three toseven ring atoms total, includes, but is not limited to oxetane,tetrahydrofuranyl, tetrahydropyranyl, aziridine, azetidine, pyrrolidine,piperidine, morpholine, piperazine and the like.

The "halogen" substituent according to the present invention may be afluoro, chloro, bromo or iodo substituent.

The language "aryl" and "heteroaryl," as used herein, refers to bothmonocyclic and polycyclic groups, which may be either substituted orunsubstituted. Examples of useful aryl ring groups include phenyl,1,2,3,4-tetrahydro-naphthyl, naphthyl, phenanthryl, anthryl, phenanthroand the like. Examples of typical heteroaryl rings include 5-memberedmonocyclic ring groups such as thienyl, pyrrolyl, imidazolyl, pyrazolyl,furyl, isothiazolyl, furazanyl, isoxazolyl, thiazolyl and the like;6-membered monocyclic groups such as pyridyl, pyrazinyl, pyrimidinyl,pyridazinyl, triazinyl and the like; and polycyclic heterocyclic ringgroups such as benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl,isobenzofuranyl, chromenyl, xanthenyl, phenoxathienyl, indolizinyl,isoindolyl, indolyl, indazolyl, purinyl, isoquinolyl, quinolyl,phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, benzothiazole,benzimidazole, tetrahydroquinoline cinnolinyl, pteridinyl, carbazolyl,beta-carbolinyl, phenanthridinyl, acridinyl, perimidinyl,phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, phenoxazinyl,and the like.

As discussed above, the R¹ substituent of formula may be hydrogen,halogen, alkyl, --OH, --O-alkyl, --O-(aryl or heteroaryl), --S-alkyl,--S-(aryl or heteroaryl), --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO,--NHOH, --NHO-alkyl, --NHNH₂, substituted --NHNH₂, --NHC(═NH)NH₂,--NHC(═NH)alkyl, fluoroalkyl, cycloalkyl, alkenyl, alkynyl, aryl, orheterocycle. The R¹ substituent is preferably a methyl or amino group.

The R² and R³ substituents of formula I according to the presentinvention, which may be the same or different, may be hydrogen, halogen,alkyl, cycloalkyl, --OH, --O-alkyl, --S-alkyl , --NH₂, --NH-alkyl,--N-(alkyl)₂, --NHCHO, --NO₂, --NHOH, --NHO-alkyl, --NHNH₂, substituted--NHNH₂, --CN, --CO₂ H, --CO₂ -alkyl, --CONH₂, --CONH-alkyl,--CON(alkyl)₂, --CSNH₂, --CSNH-alkyl, --CSN(alkyl)₂, --C(═NH)NH₂,--NHC(═NH)NH₂, --NHC(═NH)alkyl, --SO-alkyl, --SO₂ -alkyl, fluoroalkyl,--O-fluoroalkyl, --S-fluoroalkyl, --NHCO(alkyl), --NHCO(fluoroalkyl),--SO-fluoroalkyl, --SO₂ -fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂NH(alkyl), --SO₂ N(alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle.

The R² substituent is preferably hydrogen or a methyl, ethyl, hydroxy,methoxy, chloro or trifluoromethyl group. More preferably, R² ishydrogen or a methyl, chloro or trifluoromethyl group. The R³substituent is preferably hydrogen.

The Z substituent of formula I according to the present invention iseither oxygen or sulfur. In a preferred embodiment, the Z substituent isoxygen.

The R⁴ substituent of formula I according to the present invention maybe oxygen, sulfur, SO, SO₂, NH, N-alkyl, CH₂, CH-alkyl, CH-(aryl orheteroaryl), CHOH, CHO-alkyl, CHO-(aryl or heteroaryl), C(alkyl)₂,C(aryl or heteroaryl)₂, C(alkyl)(aryl or heteroaryl), CHS-alkyl,CHS-aryl, C(OH)(alkyl), C(OH)(aryl or heteroaryl), C(OH)(cycloalkyl),N(OH), N-cycloalkyl, N(cycloalkyl)SO₂, N(aryl or heteroaryl),C(cycloalkyl)₂, C(aryl or heteroaryl)(cycloalkyl), C(alkyl)(alkenyl),C(alkyl)(alkynyl), C(alkenyl)₂, C(alkynyl)₂, C(alkynyl)(aryl orheteroaryl), C(alkynyl)(alkenyl), C(alkenyl)(aryl or heteroaryl),C(cycloalkyl)(alkenyl), C(cycloalkyl)(alkynyl), C(alkyl)(aryl orheteroaryl), CH(cycloalkyl), CH(alkenyl), CH(alkynyl),C(alkyl)(cycloalkyl), C(alkyl)(O-alkyl), C(alkenyl)(O-alkyl),C(alkynyl)(O-alkyl), C(alkyl)(O-cycloalkyl), C(alkenyl)(O-cycloalkyl),C(alkynyl)(O-cycloalkyl), C(aryl or heteroaryl)(O-alkyl), C(aryl orheteroaryl)(O-cycloalkyl), C(alkynyl)(S-alkyl),C(alkynyl)(S-cycloalkyl), C(alkenyl)(S-alkyl), C(alkenyl)(S-cycloalkyl),C(alkyl)(S-alkyl), C(alkyl)(S-cycloalkyl), C(aryl orheteroaryl)(S-alkyl), C(aryl or heteroaryl)(S-cycloalkyl), N(NH₂),N[NH(alkyl)], N[N(alkyl)₂ ], N[NH(cycloalkyl)], N[N(alkyl)(cycloalkyl)],CH(NH₂), CH[NH(alkyl)], CH[NH(cycloalkyl)], CH[N(alkyl)₂ ],CH[N(alkyl)(cycloalkyl)], CH[N(cycloalkyl)₂ ], C(alkyl)(NH₂),C(alkyl)[NH(alkyl)], C(alkyl)[N(cycloalkyl)₂ ],C(alkyl)[NH(cycloalkyl)], C(alkyl)[N(alkyl)₂ ],C(alkyl)[N(alkyl)(cycloalkyl)], C(aryl or heteroaryl)(NH₂), C(aryl orheteroaryl)[NH(alkyl)], C(aryl or heteroaryl)[NH(cycloalkyl)], C(aryl orheteroaryl)[N(alkyl₂ ], C(aryl or heteroaryl)[N(cycloalkyl)₂ ], orC(aryl or heteroaryl)[N(alkyl)(cycloalkyl)].

The R⁴ substituent is preferably oxygen, sulfur or a methylene, C═O, NH,NCH₃, CH(OH) or C(OH)(phenyl) group. More preferably, the R⁴ substituentis sulfur.

The R⁵ substituent of formula I can be any one of a large number of arylor heteroaryl ring compounds, including, but not limited to, the aryland heteroaryl rings discussed previously. The R⁵ substituent may beunsubstituted or substituted. Suitable substituents for R⁵ include awide variety of electron-donating and electron-withdrawing substituents.As used herein, the language "electron-withdrawing" includes, but is notlimited to, groups such as --NO₂ ; --CF₃ ; --CN; carboxy; halogen; --SO₂R⁶, wherein R⁶ is an alkyl, aryl or heteroaryl group as discussed above,or R₆ is an --NR₇ R₈ group, wherein R₇ and R₈ represent alkyl groups;and the like. The language "electron-donating" includes, but is notlimited to, groups such as --NH₂ ; --NH-(alkyl); --NHOH; --NHNH₂ ;--O-(alkyl); --S-(alkyl); --NR⁷ R⁸, wherein R⁷ and R⁸ represent alkylgroups; and the like.

Typical substituents for R⁵ include halogen, hydroxy, alkoxy, alkyl,hydroxyalkyl, fluoroalkyl, amino, --NH-(alkyl), --N-(alkyl)₂, --CO-aminoacid, --CN, --NO₂, --CF₃, carbalkoxy, carbamyl, carbonyl, carboxy, aminoacid carbonyl, --SO₂ NHCO, SO₂ -amino acid, amino acid sulfonyl,sulfamyl, sulfanilyl, sulfhydryl, sulfino, sulfinyl, sulfo, sulfonamido,sulfonyl, (alkyl)-thio, substituted or unsubstituted phenylsulfonyl,phenylmercapto, phosphazo, phosphinico, phosphino, phospho, phosphono,phosphoro, phosphoroso, mercaptoaryl, and the like.

Particularly preferred structures for R⁵ include: ##STR2##

A preferred class of compounds according to the present inventionincludes those compounds according to formula I, wherein R³ is hydrogen.Particularly preferred compounds of this class are those wherein Z isoxygen.

Another preferred class of compounds according to the present inventionincludes those compounds according to formula I, wherein R³ is hydrogenand R¹ is either a methyl or amino group. Particularly preferredcompounds of this class are those wherein Z is oxygen.

Another preferred class of compounds according to the present inventionincludes those compounds according to formula I, wherein R³ is hydrogenand R² is hydrogen or a methyl, ethyl, hydroxy or methoxy group. Morepreferably, R² is hydrogen or a methyl group. Particularly preferredcompounds of this class are those wherein Z is oxygen.

Another preferred class of compounds according to the present inventionincludes those compounds according to formula I, wherein R³ is hydrogenand R⁴ is oxygen, sulfur or a methylene, C═O, CH(OH) or C(OH)(phenyl)group. More preferably, R⁴ is sulfur. Particularly preferred compoundsof this class are those wherein Z is oxygen.

Another preferred class of compounds according to the present inventionincludes those compounds according to formula I, wherein R³ is hydrogenand R⁵ is one of the following: ##STR3## Particularly preferredcompounds of this class are those wherein Z is oxygen.

Another preferred class of compounds according to the present inventionincludes those compounds according to formula I, wherein R³ is hydrogen,R¹ is either a methyl or amino group, R² is hydrogen or a methyl, ethyl,hydroxy or methoxy group, R⁴ is oxygen, sulfur or a methylene, C═O,CH(OH) or C(OH)(phenyl) group, and R⁵ is one of the following: ##STR4##Particularly preferred compounds of this class are those wherein Z isoxygen.

According to a preferred embodiment of the present invention, R³ ishydrogen R¹ is either a methyl or amino group, R² is hydrogen or amethyl group, R⁴ is sulfur and R⁵ is one of the rings disclosed in thepreceding paragraph. Particularly preferred compounds of this class arethose wherein Z is oxygen.

Particularly preferred compounds according to the present invention areillustrated in Table 1 below. Compounds 14A, 24A and 25A are especiallypreferred.

                                      TABLE 1                                     __________________________________________________________________________     ##STR5##                                                                     wherein R.sup.3 is H, and                                                          R.sup.1                                                                            R.sup.2                                                                             R.sup.4                                                                             R.sup.5                                                 __________________________________________________________________________     1A) CH.sub.3                                                                           H     O                                                                                    ##STR6##                                                2A) CH.sub.3                                                                           H     S                                                                                    ##STR7##                                                3A) CH.sub.3                                                                           H     CH.sub.2                                                                             ##STR8##                                                4A) CH.sub.3                                                                           H     CO                                                                                   ##STR9##                                                5A) CH.sub.3                                                                           H     CH(OH)                                                                               ##STR10##                                               6A) CH.sub.3                                                                           H     C(OH)(Ph)                                                                            ##STR11##                                               7A) CH.sub.3                                                                           H     S                                                                                    ##STR12##                                               8A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR13##                                               9A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR14##                                              10A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR15##                                              11A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR16##                                              12A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR17##                                              13A) CH.sub.3                                                                           OCH.sub.3                                                                           S                                                                                    ##STR18##                                              14A) NH.sub.2                                                                           CH.sub.3                                                                            S                                                                                    ##STR19##                                              15A) CH.sub.3                                                                           OH    S                                                                                    ##STR20##                                              16A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR21##                                              17A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR22##                                              18A) NH.sub.2                                                                           CH.sub.3                                                                            S                                                                                    ##STR23##                                              19A) CH.sub.3                                                                           CH.sub.2 CH.sub.3                                                                   S                                                                                    ##STR24##                                              20A) CH.sub.3                                                                           H     S                                                                                    ##STR25##                                              21A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR26##                                              22A) CH.sub.3                                                                           H     S                                                                                    ##STR27##                                              23A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR28##                                              24A) NH.sub.2                                                                           CH.sub.3                                                                            S                                                                                    ##STR29##                                              25A)                                                                                ##STR30##                                                               26A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR31##                                              27A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR32##                                              28A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR33##                                              29A) CH.sub.3                                                                           CH.sub.3                                                                            S                                                                                    ##STR34##                                              30A) NH.sub.2                                                                           CH.sub.3                                                                            S                                                                                    ##STR35##                                              31A) CH.sub.3                                                                           H     CH.sub.2                                                                             ##STR36##                                              32A) CH.sub.3                                                                           H     CH.sub.2                                                                             ##STR37##                                              33A) CH.sub.3                                                                           H     CH.sub.2                                                                             ##STR38##                                              __________________________________________________________________________

Another aspect of the present invention relates to processes of makingthe antiproliferative quinazoline compounds according to formula I.

One process according to the present invention for preparing quinazolinecompounds of the formula I, comprises subjecting a compound of theformula ##STR39## wherein Z and R¹ to R³ have the same meanings asdescribed previously and L is a leaving group, to a displacementreaction with the appropriate compound to cause the leaving group L tobe replaced with the desired --R⁴ --R⁵ substituent in case (i) or withthe appropriate R⁵ substituent in case (ii). The process can be carriedout under widely varying conditions, but is typically carried out in thepresence of an appropriate base, solvent and catalyst at a temperaturevarying from about 70° C. to about 165° C., preferably from about 80° C.to about 140° C., and most preferably at from about 90° C. to about 100°C.

Leaving groups suitable for use in the process described above, as wellas for use in other processes according to the present invention,include halogen atoms such as Br, Cl, F and I.

A preferred process for making antiproliferative quinazoline compoundsaccording to formula I, wherein Z and R¹ -R⁵ have the same meanings asdescribed previously, comprises the steps of:

(1) reacting a compound having the formula ##STR40## wherein L is aleaving group, for example, a halogen atom such as Br, Cl, F and I, andR² has the same meaning as described previously, with hydroxylaminehydrochloride and chloral hydrate to form an isonitrosoacetanilidecompound of the formula ##STR41## (2) treating the isonitrosoacetanilidecompound of step (1) with sulfuric acid, followed by ice andpurification with ethanol to obtain an isatin compound of the formula##STR42## (3) reacting the isatin compound of step (2) with an aqueousbasic peroxide, such as an aqueous NaOH and H₂ O₂ solution, to form ananthranilic acid compound of the formula ##STR43## (4) reacting theanthranilic acid compound of step (3) with acetic anhydride to form anacetylanthranil compound of the formula ##STR44## (5) reacting theacetylanthranil compound of step (4) with anhydrous ammonia, followed byNaOH and then by hydrochloric acid to obtain a quinazoline of theformula ##STR45## (6) subjecting the quinazoline compound of step (5) toa displacement reaction to replace the leaving group L with one of thedesired R⁴ --R⁵ substituents described previously, and thus obtain acompound according to formula I.

Step (1) can be carried out under widely varying conditions, but istypically carried out in the presence of water, chloral hydrate,hydrochloric acid, sodium sulfate and hydroxyl amine hydrochloride at atemperature varying from about 0° C. to about 100° C., preferably fromabout 20° C. to about 100° C., and most preferably at about 100° C.

Step (2) can be carried out under widely varying conditions, but istypically carried out in the presence of concentrated H₂ SO₄ at atemperature varying from about 50° C. to about 100° C., preferably fromabout 65° C. to about 100° C., and most preferably at about 80° C.

Step (3) can be carried out under widely varying conditions, but istypically carried out in the presence of water, sodium hydroxide andhydrogen peroxide at a temperature varying from about 0° C. to about 80°C., preferably from about 20° C. to about 80° C., and most preferably atabout 80° C.

Step (4) can be carried out under widely varying conditions, but istypically carried out in the presence of acetic anhydride at atemperature varying from about 70° C. to about 140° C., preferably fromabout 100° C. to about 140° C., and most preferably at about 140° C.

Step (5) can be carried out under widely varying conditions, but istypically carried out in the presence of ammonia at a temperaturevarying from about -33° C. to about 20° C., preferably at about 20° C.

Step (6) can be carried out under widely varying conditions, but istypically carried out in the presence of an appropriate base, solventand catalyst at a temperature varying from about 70° C. to about 165°C., preferably from about 80° C. to about 140° C., and most preferablyat from about 90° C. to about 100° C.

A modification to the six step process discussed above, comprises thealternate steps of:

(5a) treating the acetylanthranil compound of step (4) with MeOH,followed by hydrochloric acid, to obtain a compound of the formula:##STR46## (5a') treating the anthranilic acid of step (3) with phosgeneor triphosgene to form a compound of the formula: ##STR47## which isfurther treated with methanol; (5b) reacting the product of step (5a) or(5a') with chloroformamidine hydrochloride to obtain a quinazolinecompound of the formula: ##STR48## then subjecting the resultingquinazoline compound to a displacement reaction as set forth in step (6)discussed above to obtain a compound according to formula I.

Step (5a) can be carried out under widely varying conditions, but istypically carried out (i) in the presence of methanol at a temperaturevarying from about 0° C. to about 100° C., preferably from about 20° C.to about 70° C., and most preferably at about 70° C., and then (ii) inthe presence of concentrated hydrochloric acid at a temperature varyingfrom about 70° C. to about 100° C., more preferably at about 100° C.

Step (5a') can be carried out under widely varying conditions, but istypically carried out (i) in the presence of triphosgene at atemperature varying from about 0° C. to about 20° C., and then (ii) inthe presence of methanol at a temperature varying from about 0° C. toabout 70° C., more preferably at a temperature of from about 0° C. toabout 20° C., and most preferably at about 20° C.

Step (5b) can be carried out under widely varying conditions, but istypically carried out in the presence of diglyme and chloroformamidinehydrochloride at a temperature varying from about 100° C. to about 175°C., preferably from about 160° C. to about 175° C., and most preferablyat about 170° C.

In a particularly preferred embodiment of the six step process discussedabove, step (6) is carried out by reacting the product of either step(5) or step (5b) with a 4-thiopyridine anion, in the presence of sodiumhydride, copper (I) bromide and copper (I) oxide. A preferred processfor preparing the anions of 4-thiopyridines for use in the presentinvention comprises reacting 4-mercaptopyridine with NaH in anhydrousN,N-Dimethylacetamide. The process of preparing the 4-thiopyridines maybe carried out under widely varying conditions, but is typically carriedout in the presence of sodium hydride and dimethylformamide at atemperature varying from about -20° C. to about 20° C., preferably fromabout 0° C. to about 20° C., and most preferably at about 20° C.

Another particularly preferred process according to the presentinvention for preparing quinazoline compounds of formula I, comprisesthe steps of:

(1) reacting a compound of the formula ##STR49## where in the R⁹substituent is hydrogen, --CH₃, --OCH₃, CF₃, N(CH₃)₂, and the like, witha benzylmercaptan to form a compound of the formula ##STR50## whereinthe R¹⁰ substituent is hydrogen or --OCH₃ ; (2) reducing the product ofstep (1);

(3) deprotecting the product of step (2); and

(4) reacting the product of step (3) with a compound of the formula##STR51## wherein Z and R¹ to R³ have the same meanings as discussedpreviously and L is a leaving group, to obtain a compound of the formula##STR52##

Step (1) according to this process can be carried out under widelyvarying conditions, but is typically carried out in the presence of anappropriate base and solvent at a temperature varying from about 0° C.to about 80° C., preferably from about 0° C. to about 20° C.

Step (2), the reducing step, can be carried out under widely varyingconditions, but is typically carried out in the presence of PCl₃ andCHCl₃ at a temperature of from about 0° C. to about 80° C., preferablyfrom about 20° C. to about 80° C., and more preferably at about 20° C.

Step (3), the deprotection step, can be carried out under widely varyingconditions, but is typically carried out in the presence of anappropriate solvent and metal or metal salt at a temperature varyingfrom about -78° C. to about 20° C., preferably from about -78° C. toabout 0° C., and most preferably at from about -33° C. to about 0° C.

Step (4) can be carried out under widely varying conditions, but istypically carried out in the presence of dimethyl acetamide, sodiumhydride, copper (I) bromide and copper (I) oxide at a temperaturevarying from about 70° C. to about 165° C., preferably from about 90° C.to about 100° C., and most preferably at about 90° C.

An alternative to the four step process discussed above comprises thesteps of:

(1) reducing a compound of the formula ##STR53## to form a compound ofthe formula ##STR54## (2) reacting the product of step (1) with axanthate compound to obtain a compound of the formula ##STR55## (3)subjecting the product of step (2) to hydrolysis and further reactionwith a compound of the formula ##STR56## wherein Z and R¹ to R³ have thesame meanings discussed previously and L is a leaving group, in thepresence of N,N-Dimethylacetamide, copper (I) bromide and copper (I)oxide to obtain a compound of the formula ##STR57##

Step (1), the reducing step, can be carried out under widely varyingconditions, but is typically carried out in the presence of hydrogengas, an appropriate solvent and a catalytic amount of palladium,preferably at room temperature of about 20° C. Of course, elevatedtemperatures may be used in some cases to expedite the reaction.

Step (2) can be carried out under widely varying conditions, but istypically carried out in the presence of an aqueous acid and NaNO₂followed by potassium xanthate, at a temperature varying from about -40°C. to about 20° C., preferably from about 0° C. to about 5° C., and mostpreferably at about 0° C.

The hydrolysis part of step (3) can be carried out under widely varyingconditions, but is preferably carried out with NaOH/CH₃ OH at atemperature of from about 0° C. to about 20° C. The reaction part ofstep (3) following hydrolysis also may be carried out under widelyvarying conditions, but is typically carried out in the presence of anappropriate base, solvent and catalyst, at a temperature varying fromabout 70° C. to about 165° C., preferably from about 90° C. to about100° C., and most preferably at about 90° C.

Another preferred process for preparing the quinazoline compounds offormula I, wherein Z and R¹ to R⁵ have the same meanings as describedpreviously, comprises the steps of:

(1) reacting a compound of the formula ##STR58## wherein R¹ to R³ havethe same meanings as described previously, with a compound suitable forproviding a protecting group P, to form a compound of the formula##STR59## (2) converting the product of step (1) to a compound of theformula ##STR60## wherein L is a leaving group; (3) subjecting thequinazoline compound of step (2) to a displacement reaction to form acompound of the formula ##STR61## wherein R⁵ has the same meaning asdescribed previously; and (4) deprotecting the product of step (3).

Step (1) of the process described above can be carried out under widelyvarying conditions, but is typically carried out in the presence of theappropriate alkyl or acyl halide, a base and a solvent at a temperaturevarying from about 0° C. to about 20° C., preferably at about 20° C.

Although a variety of substituents may be used as protecting group P inthe process described above, protecting group P is preferably a CH₂ OCH₂CH₂ Si(CH₃)₃, CH₂ OCH₃, CH₂ OC(O)^(t) Bu or CO^(t) Bu group. Accordingto a preferred embodiment, P is CH₂ OCH₂ CH₂ Si(CH₃)₃.

Step (2), the converting step, can be carried out under widely varyingconditions to provide a wide variety of leaving groups, but ispreferably carried out in the presence of N-Bromosuccinimide, bromine,N-Chlorosuccinimide or N-Iodosuccinimide, at a temperature varying fromabout 20° C. to about 100° C., preferably from about 50° C. to about100° C., and most preferably at about 80° C. In a preferred embodiment,the process is carried out in the presence of N-Bromosuccinimide, CCl₄and light.

Step (3) also can be carried out under widely varying conditions, but istypically carried out in the presence of an appropriate nucleophile,base and solvent, at a temperature varying from about 0° C. to about150° C., preferably from about 20° C. to about 100° C., and mostpreferably at about 20° C.

In a preferred embodiment of this process, step (3) is carried out byreacting the product of step (2) with NaOEt (sodium ethoxide) and2-nitropropane, followed by phenylmagnesium, to form a compound of theformula ##STR62##

In another preferred embodiment, step (3) is carried out by reacting theproduct of step (2) with 5-chloroindole.

Step (4), which also can be carried out under widely varying conditions,is typically carried out in the presence of an appropriate acid or basicfluoride, at a temperature varying from about 0° C. to about 100° C.,preferably from about 20° C. to about 100° C., and most preferably atabout 20° C.

The materials and conditions used in deprotecting step (4) depend upon avariety of factors. Of course, the particular substituent used asprotecting group P is one factor. For example, when P is a CH₂ OCH₂ CH₂Si(CH₃)₃ group, step (4) is preferably carried out by reacting theproduct of step (3) with tetrabutylammonium fluoride.

In a modification of the process described above, prior to deprotectingstep (4), the product of step (3) is oxidized to form a compound of theformula ##STR63## and then, subsequent to deprotecting step (4), theproduct of step (4) is reacted with phenyllithium to form a compound ofthe formula ##STR64##

As illustrated above, it may be necessary to provide protecting groupseither before, after or during the course of preparing the compoundsaccording to the present invention.

A suitable protecting group for a ring nitrogen, such as may be includedin a heteroaryl group, is for example, a pivaloyloxymethyl group, whichmay be removed by hydrolysis with a base such as sodium hydroxide; atert-butyloxycarbonyl group, which may be removed by hydrolysis with anacid, such as hydrochloric acid or trifluoroacetic acid, or with a basesuch as tetra-n-butylammonium fluoride ("TBAF") or lithium hydroxide; amethoxymethyl group, which may be removed by hydrochloric acid andp-Toluenesulfonic acid; or a 2-(trimethylsilyl)ethoxymethyl group, whichmay be removed by TBAF or with an acid such as hydrochloric acid.

A suitable protecting group for a hydroxyl group is, for example, anesterifying group such as an acetyl or benzoyl group, which may beremoved by hydrolysis with a base such as sodium hydroxide.Alternatively, when other groups present in the starting material do notcontain an alkenyl or alkynyl group, the protecting group may be, forexample, an alpha-arylalkyl group such as a benzyl group, which may beremoved by hydrogenation in the presence of a catalyst such as palladiumon charcoal or Raney nickel. An additional protecting group for ahydroxyl group is a group such as t-butyldiphenylsilyl (--Si-t-Bu-Ph₂),which may be removed by treatment with TBAF.

A suitable protecting group for a mercapto group is, for example, anesterifying group such as an acetyl group, which may be removed byhydrolysis with a base such as sodium hydroxide.

A suitable protecting group for an amino group may be, for example, analkylcarbonyl group such as an acetyl group (CH₃ CO--), which may beremoved by treatment with an aqueous inorganic acid such as nitric,sulfuric or hydrochloric acid. Another protecting group for an aminogroup is an alkoxycarbonyl group such as a methoxycarbonyl or atert-butyloxycarbonyl group. These groups may be removed by treatmentwith an organic acid such as trifluoroacetic acid.

A suitable protecting group for a primary amino group is, for example,an acetyl group, which may be removed by treatment with an aqueousinorganic acid such as nitric, sulfuric, or hydrochloric acid, or aphthaloyl group, which may be removed by treatment with an alkylaminesuch as dimethylaminopropylamine or with hydrazine.

A suitable protecting group for a carboxy group may be an esterifyinggroup, for example, a methyl or an ethyl group, which may be removed byhydrolysis with a base such as sodium hydroxide. Another usefulprotecting group is a tert-butyl group, which may be removed bytreatment with an organic acid such as trifluoroacetic acid.

While particularly preferred processes for preparing theantiproliferative compounds according to the present invention have beendescribed in detail, it will be apparent to one skilled in the art thatvarious other processes as well as changes and modifications to thedisclosed processes can be used to prepare the compounds of the presentinvention.

The antiproliferative quinazoline compounds of the present invention,which may be employed in the pharmaceutical compositions according tothe present invention, include all of those compounds described above,as well as pharmaceutically acceptable salts of these compounds.Pharmaceutically acceptable acid addition salts of the compounds of theinvention containing a basic group are formed, where appropriate, withstrong or moderately strong organic or inorganic acids in the presenceof a basic amine by methods known in the art. Exemplary of the acidaddition salts which are included in this invention are: (1) organicacid salts such as maleate, fumarate, lactate, oxalate,methanesulfonate, ethanesulfonate, benzenesulfonate, tartrate,glucuronate, citrate, and acetate; and (2) inorganic acid salts such ashydrobromide, hydrochloride, hydrosulfate, phosphate and nitrate salts.Pharmaceutically acceptable base addition salts of compounds of theinvention containing an acidic group are prepared by known methods fromorganic and inorganic bases, and include nontoxic alkali metal andalkaline earth bases, for example, calcium, sodium and potassiumhydroxides; ammonium hydroxides; and nontoxic organic bases such astriethylamine, butylamine, piperazine andtri(hydroxymethyl)-methylamine.

As stated above, the compounds of the invention possessantiproliferative activity, a property which may express itself in theform of antitumor activity. A compound of the invention may be activeper se or it may be a pro-drug that is converted in vivo to an activecompound. Preferred compounds of the invention are active in inhibitingthe enzyme thymidylate synthase. Particularly preferred compounds areactive in inhibiting the growth of the L1210 cell line, a mouse leukemiacell line which can be grown in tissue culture. Such compounds of theinvention are also active in inhibiting the growth of bacteria such asEscherichia coli gram negative bacteria which can be grown in culture.The compounds of the invention may also be active inhibiting the growthof bacteria.

The antiproliferative compounds according to the present invention, aswell as the pharmaceutically acceptable salts thereof, may beincorporated into convenient dosage forms such as capsules, tablets, orinjectable preparations. Solid or liquid pharmaceutically acceptablecarriers may be employed. Solid carriers include starch, lactose,calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, agar,pectin, acacia, magnesium stearate and stearic acid. Liquid carriersinclude syrup, peanut oil, olive oil, saline and water.

Similarly, the carrier or diluent may include any prolonged releasematerial, such as glyceryl monostearate or glyceryl distearate, alone orwith wax. When a liquid carrier is used, the preparation may be in theform of a syrup, elixir, emulsion, soft gelatin capsule, sterileinjectable liquid (e.g. solution), such as an ampoule, or an aqueous ornonaqueous liquid suspension.

The pharmaceutical preparations are made following conventionaltechniques of a pharmaceutical chemist involving steps such as mixing,granulating and compressing, when necessary for tablet forms; or mixing,filling and dissolving the ingredients, as appropriate, to give thedesired products for oral, parenteral, topical, intravaginal,intranasal, intrabronchial, intraoccular, intraaural and rectaladministration.

The composition of the invention may further comprise one or more othercompounds which are antitumor agents, such as a mitotic inhibitors(e.g., vinblastine), alkylating agents (e.g., cisplatin, carboplatin andcyclophosphamide), dihydrofolate reductase inhibitors (e.g.,methotrexate, piritrexim and trimetrexate), other thymidylate synthaseinhibitors, antimetabolites (e.g., 5-fluorouracil and cytosinearabinoside), intercalating antibiotics (e.g., adriamycin andbleomycin), enzymes (e.g., asparaginase), topoisomerase inhibitors(e.g., etoposide) or biological response modifiers (e.g., interferon).

The composition of the invention may also comprise one or more othercompounds, including antibacterial, antifungal, antiparasitic,antiviral, antipsoriatic and anticoccidial agents. Exemplaryantibacterial agents include, for example, sulfonamide such assulfamethoxazole, sulfadiazine, sulfameter or sulfadoxine; dihydrofolatereductase inhibitors such as trimethoprim, bromodiaprim or trimetrexate;penicillins; cephalosporins; aminoglycosides; bacteriostatic inhibitorsof protein synthesis; the quinolonecarboxylic acids and their fusedisothiazolo analogs.

Another aspect of the invention relates to a therapeutic process ofinhibiting the growth and proliferation of cells of higher organisms andmicroorganisms, which process comprises administering to a host, such asa vertebrate host (e.g., a mammal or bird) an effective amount of acompound according to the present invention. A particularly preferredtherapeutic process comprises administering to a host an effectiveamount of a compound according to the present invention to inhibit theenzyme thymidylate synthase, The compounds of the invention areparticularly useful in the treatment of mammalian hosts, such as humanhosts, and in the treatment of arian hosts.

Any of the antiproliferative compounds described above, orpharmaceutically acceptable salts thereof, may be employed in thetherapeutic process of the invention. The compounds of the invention maybe administered in the form of a pharmaceutically acceptable compositioncomprising a diluent or carrier, such as those described above. Doses ofthe compounds preferably include pharmaceutical dosage units comprisingan efficacious quantity of active compound. By an efficacious quantityis meant a quantity sufficient to inhibit the folate metabolic pathwaysand derive the beneficial effects therefrom through administration ofone or more of the pharmaceutical dosage units. An exemplary dailydosage unit for a vertebrate host comprises an amount of up to about 1gram of active compound per kilogram of the host, preferably one half ofa gram, more preferably 100 milligrams, and most preferably about 50milligrams per kilogram of the host.

The selected dose may be administered to a warmblooded animal or mammal,for example a human patient, in need of treatment mediated by folatemetabolic pathways inhibition by any known method of administration,including topically (e.g. as an ointment or cream), orally, rectally(e.g., as a suppository), parentally, by injection or continuously byinfusion, intravaginally, intranasally, intrabronchially, intraaurallyor intraocularly.

The compounds according to the present invention may be characterized asproducing any one or more of an antiproliferative effect, anantibacterial effect, an antiparasitic effect, an antiviral effect, anantipsoriatic effect, an antiprotozoal effect, an anticoccidial effector an antifungal effect. The compounds are especially useful inproducing an antitumor effect in a vertebrate host harboring a tumor.

EXAMPLES

As stated previously, Table 1 discloses a number of preferred compoundsaccording to the present invention. Examples of the process used to makeseveral of these preferred compounds is set forth below.

The structures of all compounds of the invention were confirmed byproton magnetic resonance spectroscopy, infrared spectroscopy, elementalmicroanalysis and/or mass spectrometry. Infrared absorption spectra weretaken on a Midac FT or a Perkin Elmer Model 457 spectrophotometer.Spectra were obtained as KBr (potassium bromide) pellets or neat films,and the peak values were reported in cm⁻¹.

Proton magnetic resonance spectra were determined using a GeneralElectric QE-300 spectrometer operating at a field strength of 300 MHz.Chemical shifts are reported in parts per million (δ) by setting thereferences such that, in CDCl₃, the CDCl₃ peak is at 7.26 ppm and, inDMSO-d₆, the DMSO peak is at 2.49 ppm. Standard and peak multiplicitiesare designated as follows: s, singlet; d, doublet; dd, doublet ofdoublets; t, triplet; brs, broad singlet; brd, broad doublet; br, broadsignal; m, multiplet.

Mass spectra were determined using a VG 7070E-HF high resolution massspectrometer using the direct insertion method, an ionizing voltage of70 eV, and an ion source temperature of 200° C. Elemental microanalysisprovided results for the elements usually within ±0.4% of thetheoretical values.

General Procedures

N-N-Dimethylformamide ("DMF") was dried over activated (250° C.) 3-Åmolecular sieves; N,N-dimethylacetamide ("DMA") (Aldrich Gold Labelgrade) was similarly dried. Tetrahydrofuran ("THF") was distilled fromsodium benzophenone ketyl under nitrogen. The term "ether" refers todiethyl ether.

Flash chromatography was performed using Silica gel 60 (Merck Art 9385).Where the crude solid was insoluble in the chosen eluant, it wasdissolved in a more polar solvent, and Merck Art 7734 silica was added.The slurry was evaporated to dryness on a rotary evaporator fitted witha course glass frit to prevent spraying of the silica. The coated silicawas then applied to the column. Thin layer chromatographs ("TLC") wereperformed on precoated sheets of silica 60 F₂₅₄ (Merck Art 5719).Extracts were dried over anhydrous Na₂ SO₄ or MgSO₄. Melting points weredetermined on a Mel-Temp apparatus and were uncorrected.

Example 1

Preparation of Compounds 8A and 14A

Compounds 8A and 14A were prepared according to the following reactionscheme: ##STR65## Preparation of Intermediate Compound(1)--3-Bromo-4-methyl-aniline

A solution of 50.0 g (0.23 mol) 2-Bromo-4-nitro-toluene in 500 mlmethanol was placed in a Parr hydrogenation bottle. To the solution wasadded 5.0 g Raney nickel. This mixture was hydrogenated at 30 psi H₂ onthe Parr hydrogenator for three hours with agitation. The Parr bottlewas vented, the reaction mixture was filtered through diatomaceous earth(celite), and the filtrate was evaporated to yield 41.0 g (95%) of ayellow oil. IR (neat) 3329, 3144, 2604, 1609, 1288, 1030, 812 cm⁻¹ ; ¹ HNMR (DMSO-d₆) δ2.13 (s, 3H), 5.60 (bs, 2H), 6.46 (dd, 1H, J=8.1 Hz, 2.3Hz), 6.79 (d, 1H, J=2.3 Hz), 6.94 (d, 1H, J=8.2 Hz). HRMS calcd. for C₇H₈ BrN: 184.9843. Found: 184.9840.

Preparation of Intermediate Compound(2)--3-Bromo-4-methyl-α-isonitrosoacetanilide

A mixture of 45.0 g chloral hydrate (0.27 mol), 65.0 g sodium sulfate(0.46 mol), 40.0 g 3-Bromo-4-methyl-aniline (1) (0.21 mol), 20 mlconcentrated HCl, 55.0 g of hydroxylamine hydrochloride (0.79 mol) and1.5 l of H₂ O were heated at 100° C. for one hour. The reaction mixturewas cooled to 0° C., and the precipitate was collected by filtration.The solid was washed with H₂ O and dried to yield 41.0 g (76%) as a tansolid: M.P. 195°-197° C. IR (KBr) 3439, 3310, 3110, 2998, 2876, 2749,1636, 1591, 1466, 1256, 905, 691 cm⁻¹ ; ¹ H NMR (DMSO-d₆) δ2.28 (s, 3H),3.50 (bs, 1H), 7.28 (d, 1H, J=8.3 Hz), 7.53 (dd, 1H, J=8.2, 2.1 Hz),8.02 (d, 1H, J=2.0 Hz), 10.26 (s, 1H), 12.21 (s, 1H). Anal. Calcd. forC₉ H₉ BrN₂ O₂ : C, 42.04; H, 3.53; Br, 31.08; N, 10.90. Found: C, 42.71;H, 3.57; Br, 31.44; N, 11.09.

Preparation of Intermediate Compound (3)--4-Bromo-5-methylisatin

To 160 ml concentrated sulfuric acid at 80° C. was added 40 g (0.156mol) of (2) and stirred for one hour. The reaction mixture was cooled toroom temperature and then poured onto 2 l of crushed ice. Theprecipitate was filtered, washed with water and then washed withbenzene. The red solid was added to 800 ml of boiling ethanol. Thesolution was allowed to cool to room temperature, collected and thenwashed with cold ethanol. 6-Bromo-5-methylisatin, as well as some of thedesired product remains in the mother liquor, and can be separated bysilica gel flash column chromatography. The filter cake was dried toyield 19 g (50.7%) of a red solid: M.P. 245°-248° C. IR (KBr) 3302,1750, 1609, 1466, 1273, 675 cm⁻¹ ; ¹ H NMR (CDCl₃) δ2.26 (s, 3H), 6.8(d, 1H, J=7.9 Hz), 7.5 (d, 1H, J=8.3 Hz), 11.06 (s, 1H). Anal. Calcd.for C₉ H₆ BrNO₂ : C, 45.02; H, 2.52; Br, 33.28; N, 5.86. Found: C,45.10; H, 2.54; Br, 33.19; N, 5.84.

Preparation of Intermediate Compound (4)--5-Methyl-6-bromo-anthranilicacid

A mixture of 80 ml 3N NaOH and 19 g of isatin (3) (0.08 mol) were heatedat 80° C. To the solution was added 18 ml 30% H₂ O₂, and the mixture wasstirred for one hour. The mixture was cooled to 5° C. and acidified topH5 with concentrated hydrochloric acid. The solution was evaporated todryness and then added to 300 ml methanol. The mixture was filtered, andthe filtrate was evaporated to yield 18 g of a tan solid (97.8% theory):M.P. (hydrochloride) 290°-294° C. IR (KBr) 3619, 3229, 1578, 1478, 1412,1381, 1084, 1010, 820, 706 cm⁻¹ ; ¹ H NMR (DMSO-d₆) δ2.13 (s, 3H), 4.9(s, 2H), 6.4 (d, 1H, J=7.9 Hz), 6.74 (d, 1H, J=7.8 Hz).

Preparation of Intermediate Compound (5)--5-Bromo-6-methylacetylanthranil (5-bromo-2,6-dimethyl-4H-3,1-benzoxazin-4-one)

A mixture of 18 g anthranilic acid (4) (0.078 mol) in 300 ml aceticanhydride was heated at reflux for 3 hours. The solution was cooled to0° C. and filtered. The filter cake was washed with acetone to yield 16g (81% theory) as a white solid (M.P. 190°-194° C.) which was usedwithout further purification. IR (KBr) 3460, 1750, 1660, 1574, 1416,1260, 1070, 841 cm⁻¹ ; ¹ H NMR (CDCl₃) δ2.45 (s, 3H), 2.55 (s, 3H), 7.40(d, 1H, J=8.2 Hz), 7.64 (d, 1H, J=8.0 Hz). HRMS calcd. for C₁₀ H₈ BrNO₂: 252.9738. Found: 252.9743.

Preparation of Intermediate Compound(6)--5-Bromo-3,4-dihydro-2,6-dimethylquinazolin-4-one

Anhydrous ammonia (50 ml) was condensed into a flask containing 8.5 g(34.0 mmol ) anthranil (5), and the reaction was stirred for 3 hrs. Thesolvent was evaporated to give a residue, and 75 ml of 1N NaOH wasadded. The reaction mixture was heated at reflux temperature for 1 hr.The resulting solution was cooled to 0° C. and acidified to pH4 withconcentrated hydrochloric acid. The mixture was filtered, and the filtercake was washed with water and then dried to yield 7.1 g (82.5% theory)of 6 as a tan solid: M.P. 288°-291° C. (dec.) The product was usedwithout further purification. IR (KBr) 2910, 2620, 1680, 1630, 1460,1377, 1298, 1128, 872 cm⁻¹ ; ¹ H NMR (DMSO-d₆) δ2.33 (s, 3H), 2.43 (s,3H), 7.49 (d, 1H, J=8.3 Hz), 7.70 (d, 1H, J=8.3 Hz), 12.20 (bs, 1H).HRMS calcd. for C₁₀ H₉ BrN₂ O: 251.9898. Found: 251.9908.

Preparation of Compound (7)--Methyl-2-amino-6-bromo-5-methylbenzoate

A mixture of 10 g (0.039 mol) anthranil (5) in 75 ml methanol was heatedat reflux for 2 hrs. To the solution was added 10 ml concentratedhydrochloric acid, and the mix was heated for an additional two hours.The reaction mixture was evaporated to dryness. The residue wasdissolved in 20 ml H₂ O and neutralized to pH7 with triethylamine. Theaqueous solution was extracted with methylene chloride. The layers wereseparated, and the organic layer was dried over magnesium sulfate,filtered and evaporated to dryness to yield 6.0 g of (7), as an orangeoil (63% theory). IR (neat) 3483, 3410, 3220, 3000, 2950, 2851, 1720,1620, 1560, 1430, 1288, 1120, 1015, 816 cm⁻¹ ; ¹ H NMR (CDCl₃) δ2.31 (s,3H), 3.95 (s, 3H), 4.10 (bs, 2H), 6.60 (d, 1H, J=8.2 Hz), 7.05 (d, 1H,J=8.1 Hz). HRMS calcd. for C₉ H₁₀ BrNO₂ : 242.9890. Found: 242.9895.

Preparation of Intermediate Compound(8)--2-Amino-5-bromo-3,4-di-hydro-6-methylquinazolin-4-one

To a solution of methyl ester (7) (6 g, 24 mmol) in 50 ml of diglyme wasadded 3 g (24 mmol) of chloroformamidine hydrochloride. The mixture washeated at reflux for 1 hr. The mixture was cooled to 0° C. and filtered.The solid was washed with ether and then dried to yield 6.25 g (88%theory) of a tan solid: M.P. (hydrochloride) >390° C. The product wasused without further purification. IR (KBr) 3140, 2950, 1670, 1620,1471, 1402, 816, 600 cm⁻¹ ; ¹ H NMR (DMSO-d6) δ2.28 (s, 3H), 6.75 (bs,2H), 7.0 (d, 1H, J=8.3 Hz), 7.40 (d, 1H, J=8.0 Hz), 11.8 (bs, 1H). HRMScalcd. for C₉ H₈ BrN₃ O: 253.9927. Found: 253.9929.

Preparation of Compound (9) (Compound8A)--3,4-Dihydro-2,6-dimethyl-4-oxo-5-(4-pyridylthio)-quinazoline

To a solution of 3.2 g 4-mercaptopyridine (28.8 mmol) in ml of anhydrousN,N-Dimethylacetamide at 0° C. was added 1.24 g (28.8 mmol) NaH (60%dispersion in mineral oil), and the mix was stirred for 1 hr. To thisreaction mixture was added 3.1 g bromoquinazoline (6) (0.012 mol), 1.4 gcopper (I) bromide, and 0.70 g of copper (I) oxide. The mix was heatedat 90° C. for 4 hrs. The reaction mixture was evaporated to dryness, 50ml of an H₂ S/methanol solution (10 g/l) was added to the residue, andthe mixture was stirred for 1 hr. The mixture was filtered, and thefiltrate was evaporated to dryness. The solid was purified via flashchromatography on silica gel using MeOH/CH₂ Cl₂ (5:95) to yield 1.7 g(48% theory) of a tan solid: M.P. 235°-238° C.; IR (KBr) 3430, 1670,1633, 1575, 1460, 1408, 1300, 841, 820, 714 cm⁻¹ ; ¹ H NMR (DMSO-d₆)δ2.28 (s, 3H), 2.40 (s, 3H), 6.80 (d, 2H, J=5.9 Hz), 7.60 (d, 1H, J=8.3Hz), 7.80 (d, 1H, J=8.5 Hz), 8.24 (d, 2H, J=6.5 Hz), 12.10 (bs, 1H).Anal. Calcd. for C₁₅ H₁₃ N₃ OS.H₂ O: C, 59.80; H, 4.98; N, 13.95; S,10.63. Found: C, 59.58; H, 4.90; N, 13.89; S, 10.62. HRMS Calcd. for C₁₅H₁₃ N₃ OS: 283.0773. Found: 283.0779.

Preparation of Compound (10) (Compound14A)--2-Amino-3,4-dihydro-6-methyl-4-oxo-5-(4-pyridylthio)-quinazoline

To a solution of 17.2 g 4-mercaptopyridine (15.5 mol) in 250 ml ofanhydrous N,N-Dimethylacetamide at 0° C. was added 6.2 g of (15.5 mol)NaH (60% dispersion in mineral oil), and the reaction was stirred for 1hr. To the solution was added 15 g aminoquinazoline.HCl (8) (51.3 mmol),4.5 g copper (I) bromide, and 4.5 g copper (I) oxide. The mixture washeated at 90° C. for 4 hrs., and then concentrated under vacuum. To theresulting solid was added 150 ml H₂ S/MeOH solution (20 g/l). The darkmixture was stirred for 1 hr., the precipitated CuS was removed byfiltration, and the methanolic filtrate was evaporated. The solid waswashed with methylene chloride, followed by ethyl ether and finallyboiling isopropanol to yield 7.5 g (50% theory) of (10) as a tan solid:M.P. 301°-302° C.; IR (KBr) 3320, 3150, 2750, 1670, 1575, 1466, 1305,1220, 804, 710, 482 cm⁻¹ ; ¹ H NMR (DMSO-d₆) δ2.30 (s, 3H), 6.35 (bs,2H), 6.80 (d, 2H, J=5.9 Hz), 7.26 (d, 1H, J=8.4 Hz), 7.58 (d, 1H, J=8.5Hz), 8.25 (bs, 2H), 10.85 (bs, 1H). Anal. Calcd. for C₁₄ H₁₂ N₄ OS.1.5H₂ O: C, 54.00; H, 4.86; N, 18.00; S, 10.30. Found: C, 53.81; H, 4.25;N, 17.71; S, 10.28. HRMS calcd. for C₁₄ H₁₂ N₄ OS: 284.0734. Found:284.0732.

Example 2

Preparation of Compounds 13A and 15A

Compounds 13A and 15A were prepared according to the following reactionscheme: ##STR66## Preparation of Intermediate Compound(11)--3-Bromo-4-methoxyaniline

To a solution of 38.0 g of 1-bromo-4-nitroanisole (0.164 mol) in 300 mlof methanol/THF (1:1) was added 5 ml of anhydrous hydrazine and 4.0 g ofactivated Raney nickel catalyst suspended in ethanol. The mixture wasstirred and heated to a gentle reflux, where upon the mixture began toeffervesce Within a period of 3 hrs., 7 additional ml of hydrazine and 4additional grams of Raney nickel were introduced into the reaction. Thewarm reaction mixture was vacuum filtered through a pad of silica gel toremove the catalyst, and the pad was thoroughly washed with ethylacetate. The filtrate was concentrated, and the dark brown oil wasplaced under high vacuum to remove traces of solvent. The productdecomposes readily and was used as is. ¹ H NMR (CDCl₃) δ3.46 (s, 2H),6.60 (dd, 1H, J=8.6, 2.7 Hz), 6.73 (d, 1H, J=8.6 Hz), 6.92 (d, 1H, J=2.7Hz).

Preparation of Intermediate Compound(12)--3-Bromo-4-methoxy-α-isonitrosoacetanilide

In a 250 ml 3-neck round bottom flask, 84 ml water was added to 6.3 g(37.8 mmol) chloralhydrate. The flask was fitted with a mechanicalstirrer and reflux condenser, and 90 g anhydrous sodium sulfate powderwas added over a period of 1 minute with constant stirring. A solutionof 6.3 g (31.2 mmol) of-aniline (11) in 3.0 ml conc. HCl and 21 ml waterwas added, followed by a solution of 7.7 g (112 mmol H₂ NOH.HCl in 35 mlwater. The mixture was slowly heated to reflux with constant stirringand continued for 2 minutes at which time brown crystals formed. Themixture was cooled, the solid filtered off, washed well with water anddried to constant weight by vacuum. The resulting solid weighed 5.65 g(66% theoretical) and was pure enough for the next step. An analyticalsample was prepared by recrystallization. M.P. 202°-203° C.(hexane,EtOAc). IR (KBr) 3409, 2875, 2056, 2023, 1643, 1634, 1543, 1502, 1295,1270, 1047, 799 cm⁻¹ ; ¹ H NMR (CDCl₃, one drop DMSO-d₆) δ3.88 (s, 3H),6.87 (d, 1H, J=8.9 Hz), 7.53 (m, 2H), 7.83 (d, 1H, J=2.5 Hz), 8.49 (s,1H), 11.60 (s, 1H, NH). Anal. Calcd. for C₉ H₉ BrN₂ O₃.0.11 EtOAc: C,40.09; H, 3.52; Br, 28.26; N, 9.91. Found: C, 40.45; H, 3.44; Br, 27.86;N, 10.34.

Preparation of Intermediate Compound (13)--4-Bromo-5-methoxyisatin

Vacuum dried α-Isonitrosoacetanilide (12) (3.0 g; 11 mmol) was slowlyadded to 8 ml conc. H₂ SO₄ at 50° C. while being stirred. The reactionmixture first became yellow, and then turned dark. The temperature wasraised to 65° C. for 10 minutes, and the reaction was followed byTLC(EtOAc/hexane; 40:60). Heating at 65°-70° C. was resumed until allthe starting material was consumed as judged by TLC. Upon completion,the reaction mixture was cooled and added to 80 g of crushed ice withstirring. A dark red solid formed and was filtered off, washed free ofacid by water and dried under vacuum. The resulting substance waspurified by chromatography on a flash silica column using a gradientsystem of EtOAc/hexane; 40:60; 50:50; 60:40; 70:30; 80:20. The undesiredisomer, 6-Bromo-5-methoxyisatin eluted first, followed by the desiredisomer (13), which was isolated as a red solid (0.71 g; 25% yield). M.P.250°-251° C. IR (KBr) 2064, 1758, 1750, 1634, 1278 cm⁻¹ ; ¹ H NMR(CDCl₃, one drop DMSO-d₆) δ3.91 (s, 3H), 6.84 (d, 1H, J=8.8 Hz), 7.09(d, 1H, J=8.8 Hz), 10.88 (s, 1H). Anal. Calcd. for C₉ H₆ BrNO₃ : C,42.19; H, 2.34; Br, 31.25; N, 5.47. Found: C, 42.27; H, 2.37; Br, 31.30;N, 5.42.

Preparation of Intermediate Compound(15)--5-Bromo-6-methoxyacetylanthranil(5-bromo-2,6-dimethyl-4H-3,1-benzoxazin-4-one)

A magnetically stirred solution of 2.28 g (8.9 mmol) isatin (13), in13.4 ml of 2N aq. NaOH (26.7 mmol) was cooled to 0° C. To this coldsolution 0.90 ml of 30% H₂ O₂ (8.9 mmol) was added gradually keeping thetemperature below 20° C. The progress of the reaction was followed byTLC(EtOAc/hexane; 40:60). An additional 0.20 ml of 30% H₂ O.sub. wasadded, and the reaction mixture was stirred 20 minutes at roomtemperature. At this time, TLC indicated consumption of the startingmaterial. The mixture was acidified with glacial acetic acid to pH4 andconcentrated via a cryogenic trap at -78° C., leaving crude6-Bromo-5-methoxyanthranilic acid (14), as a grey semi-solid. Thisslurry was treated with 28 ml of acetic anhydride and refluxed for 40minutes. The dark mixture was then concentrated as before. To theresidue was added an excess of ethyl acetate:hexane (2:1). The mixturewas heated and filtered hot through silica gel to remove insoluble andcolored particulants. The solution was partially concentrated andallowed to cool, and the product crystallized yielding 1.71 g (71% basedon starting isatin (13)). M.P. 228°-229° C. (dec.). IR (KBr) 3397, 2039,1717, 1651, 1625, 1543, 1295, 1055, 881, 617 cm⁻¹ ; ¹ H NMR (CDCl₃)δ2.42 (s, 3H), 3.98 (s, 3H), 7.34 (d, 1H, J=8.9 Hz), 7.51 (d, 1H, J=8.9Hz), Anal. Calcd. for C₁₀ H₈ BrNO₃ : C, 44.44; H, 2.96; Br, 29.62; N,5.19. Found: C, 44.32; H, 3.04; Br, 29.53; N, 5.09.

Preparation of Intermediate Compound(16)--5-Bromo-3,4-dihydro-6-methoxy-2-methyl-quinazolin-4-one

To 1.25 g (4.6 mmol) of the anthranil (15), in a dried round bottomflask equipped with a dry ice condenser, was condensed approximately 50ml of anhydrous NH₃. The mixture was magnetically stirred for 40 min. Atthis time, the dry ice condenser was removed, and the NH₃ was allowed toevaporate. Upon evaporation, 15 ml water and 1.5 ml of 2N NaOH wereadded, and the solution was refluxed for 1 hr. The solution was thencooled to room temperature, and 1N HCl was added adjusting the pH toapproximately 9 and thus precipitating the quinazoline. The whitesubstance was filtered off, washed with water and dried, yielding 0.71 g(57%). M.P. 273°-274° C. IR (KBr) 3189, 3074, 2990, 2974, 2899, 2362,1676, 1643, 1552, 1461, 1303, 1286, 1063, 872, 832 cm⁻¹ ; ¹ H NMR(CDCl₃) δ2.39 (s, 3H), 3.98 (s, 3H), 7.39 (d, 1H, J=9.0 Hz), 7.59 (d,1H, J=9.0 Hz), 11.60 (s, 1H), Anal. Calcd. for C₁₀ H₉ BrN₂ O₂ : C,44.61; H, 3.35; Br, 29.74; N, 10.41; Found: C, 44.56; H, 3.40; Br,29.63; N, 10.36.

Preparation of Intermediate Compound (17) (Compound13A)--3,4-Dihydro-6-methoxy-2-methyl-4-oxo-5-(4-pyridylthio)-quinazoline

To 78 mg (0.7 mmol) of 4-Mercaptopyridine was added 34 mg (0.5 mmol) ofsolid NaOH in 1 ml of dry DNA. To the resulting solution, 134 mg (0.5mmol) of quinazolinone (16) dissolved in 2 ml dry DMA was added. Themixture was kept under N₂, and a finely ground catalyst mixturecontaining 44 mg CuBr and 22 mg Cu₂ O was added The mixture was stirredmagnetically and heated to 135° C., until the reaction was complete asJudged by TLC (anh. NH₃ /MeOH/CHCl₃ ; 0.5: 4.5: 9.5). The solvent wasremoved under high vacuum through a cryogenic trap cooled to -78° C. Thedesired product was isolated by flash chromatography (anh. NH₃/MeOH/CHCl₃ ; 0.5: 4.5: 9.5) on silica, yielding 130 mg (89%) of (17) asa white powder. M.P. 248°-249° C. (dec.). IR (KBr) 3358, 3073, 2933,1682, 1634, 1574, 1475, 1462, 1318, 1277, 1059, 835, 710 cm⁻¹ ; ¹ H NMR(CDCl₃) δ2.36 (s, 3H), 3.84 (s, 3H), 6.90 (d, 2H, J=5.1 Hz), 7.48 (d,1H, J=9.1 Hz), 7.79 (d, 1H, J=9.1 Hz), 8.28 (d, 2H, J=5.1 Hz), 10.86 (s,1H). Anal. Calcd. for C₁₅ H₁₃ N₃ O₂ S: C, 60.18; H, 4.38; N, 14.04; S,10.71. Found: C, 60.28; H, 4.43; N, 14.07; S, 10.63. HRMS Calcd. for C₁₅H₁₃ N₃ O₂ S: 299.0730. Found: 299.0718.

Preparation of Intermediate Compound (18) (Compound15A)--3,4-Dihydro-6-hydroxy-2-methyl-4-oxo-5-(4-pyridylthio)-quinazoline

To cleave the methyl ether, quinazoline (17) (100 mg; 0.30 mmol) wasgently refluxed with 2 ml of a 1:1 mixture of 48% aq. HBr and glacialAcOH for 8 hrs. At this time, the solvent was removed via high vacuumthrough a cryogenic trap at -78° C. The obtained residue was dissolvedin 10% anh. NH₃ in MeOH, and subjected to flash column chromatography onsilica (anh. NH₃ /MeOH/CHCl₃ ; 0.5: 4.5: 9.5) yielding 62 mg of (18) asa white powder (65%) M.P. 246°-247° (dec). IR (KBr) 3450, 3240, 3073,1667, 1634, 1580, 1464, 629 cm⁻¹ ; ¹ H NMR (CDCl₃) δ2.40 (s, 3H), 6.83(d, 2H, J=6.0 Hz), 7.40 (d, 1H, J=9.0 Hz), 7.59 (d, 1H, J=9.0 Hz), 8.20(d, 2H, J= 6.0 Hz), 8.51 (s, 1H), 11.51 (s, 1H). Anal. Calcd. for C₁₄H₁₁ N₃ O₂ S: C, 58.94; H, 3.13; N, 11.97; S, 11.22. Found: C, 58.98; H,3.16; N, 12.00; S, 11.61. HRMS calcd. for C₁₄ H₁₁ N₃ O₂ S: 285.05733.Found: 285.05720.

Example 3

Preparation of Compound 12A

Compound 12A was prepared according to the following reaction scheme:##STR67## Preparation of Compound (19)--4-Benzylthio-2-picoline-N-oxide

Mineral oil was removed from potassium hydride (0.11M; 35 wt. %dispersion in mineral oil) by several washings with petroleum ether(5×50 ml). The remaining petroleum ether was removed under vacuum. Tothis dry solid, 350 ml of anhydrous THF was added cautiously. The wellstirred suspension was cooled to 0° C. To this mixture, 14.1 ml (0.12mol) of benzylmercaptan was added dropwise over a period of 30 minutes.The resultant milky white mixture was warmed to room temperature andallowed to stir for an additional 30 minutes The mixture was then cooledto -30° C., and 15.41 g (0.1 mol) of 4-nitro-2-picoline-N-oxide wasadded portionwise. The mixture became dark orange-brown in color. Oncewarmed to room temperature, the mixture was refluxed for one hour. Atthis time, the reaction was cooled to 0° C. and quenched with 50 ml ofwater. The pH of the mixture was adjusted to approximately 6 with 2MHCl, and extracted with dichloromethane (3×300 ml). The combined organiclayers were dried (anhydrous Na₂ SO₄), and the solvent was removed underreduced pressure. The crude residue was chromatographed on flash silicagel with MeOH/CH₂ Cl₂ (Gradient: 3:97, 4:96, 5:95). The pure product wasisolated (6.94 g; 30% yield) as a tan solid: M.P. 98°-99° C.; IR (KBr)3063, 3028, 1612, 1466, 1236, 831, 715, 675 cm⁻¹ ; ¹ H NMR (CDCl₃) δ2.45(s, 3H), 4.16 (s, 2H), 6.97 (dd, 1H, J=6.8, 2.7 Hz), 7.07 (d, 1H, J=2.7Hz), 7.32 (m, 5H), 8.09 (d, 1H, J=6.8 Hz). Anal. Calcd. for C₁₃ H₁₃ NOS:C, 67.50; H, 5.66; N, 6.05; S, 13.86. Found: C, 67.51; H, 5.69; N, 6.08;S, 13.77.

Preparation of Intermediate Compound (20)--4-Benzylthio-2-picoline

Compound (19), (1.97 g, 8.5 mmol) was dissolved into 50 ml ofchloroform. The solution was stirred, cooled to 0° C., and 1.75 ml (17.4mmol) of phosphorous trichloride was added dropwise. Once the additionwas complete, the reaction mixture was brought to room temperature andthen heated slightly under reflux temperature (approximately 55° C.)until no starting N-oxide was present by TLC (MeOH/CH₂ Cl₂ ; 5:95). Thesolution was then recooled to 0° C., and 10 gm of ice was added withvigorous stirring. The mixture was made basic (pH 8) by careful additionof 1M NaOH, and the organic phase was separated. The aqueous layer wasextracted with dichloromethane (3×50 ml), and the organic layers werecombined and dried (anhydrous Na₂ SO₄). Removal of the solvent underreduced pressure gave an oil which was chromatographed on a short flashsilica column using MeOH/CH₂ Cl₂ ; 3:97. The product was isolated as awhite solid (1.54 g; 84% yield): M.P. 69°-70° C.; IR (KBr) 3028, 3003,2920, 1583, 1454, 864, 815, 719, 702 cm⁻¹ ; ¹ H NMR (CDCl₃) δ2.55 (s,3H), 4.22 (s, 2H), 7.03 (m, 2H), 7.35 (m, 5H), 8.28 (d, 1H, J=5.5 Hz).Anal. Calcd. for C₁₃ H₁₃ NS: C, 72.52; H, 6.08; N, 6.50; S, 14.90.Found: C, 72.46; H, 6.11; N, 6.50; S, 14.80

Preparation of Compound (21) (Compound12A)--3,4-Dihydro-2,6-dimethyl-4-oxo-5-[4-(2-picolinylthio)]-quinazoline

To a solution of 5 ml NH₃ condensed into 5 ml THF kept at -78° C. wasadded 115 mg sodium metal (5.0 mmol). The deep blue solution was stirredfor 15 minutes. To the reaction mixture was added 1.0 g (4.65 mmol) of4-benzylthio-2-picoline (20), and the reaction was stirred for 11/2 hrs.at 0° C. The solvent was removed under vacuum, and to the resultingsolid was added 10 ml of anhydrous N,N-Dimethylacetamide, 0.5 gquinazoline (6) (2.0 mmol) and 0.25 g of copper (I) bromide. The mix washeated at 90° C. for 4 hrs. The solvent was removed under vacuum, andthe solid was treated with 10 ml of H₂ S/MeOH solution (20 g/l). Theinsoluble CuS was filtered off, and the filtrate was evaporated todryness. The solid was purified using flash chromatography on silicawith MeOH/CH₂ Cl (5:95) to yield 400 mg (84% theory) of a tan solid:M.P. 225°-227° C.; IR (KBr) 3480, 3160, 3053, 2960, 1670, 1630, 1590,1460, 1298, 831 cm⁻¹ ; ¹ H NMR (DMSO-d₆) δ2.28 (s, 6H), 2.36 (s, 3H),6.60 (bs, 1H), 6.80 (6s, 1H), 7.60 (d, 1H, J=8.4 Hz), 7.80 (d, 1H, J=8.4Hz). Anal. Calcd. for C₁₆ H₁₅ N₃ OS.0.5 H₂ O: C, 62.73; H, 5.22; N,13.72; S, 10.46. Found: C, 63.08; H, 5,20; N, 13.73; S, 10.50. HRMSCalcd. for C₁₆ H₁₅ N₃ OS: 297.0936. Found: 297.0936.

Example 4

Preparation of Compound 16A

Compound 16A was prepared according to the following reaction scheme:##STR68## Preparation of Intermediate Compound(22)--2-Methoxypyridine-N-oxide

This compound, originally prepared by H. J. Den Hertog and M. VanAmmers, Rec. Trav. Chim. 1955, 74, 1160, was synthesized using adifferent procedure. To a solution of 21.83 of 2-methoxypyridine (0.2mol) in glacial acetic acid (80 ml), was cautiously added 30% hydrogenperoxide (20 ml). The stirred mixture was heated to 80° C. for 3 hrs,and cooled to room temperature. An additional 20 ml of 30% H₂ O₂ wasadded, and the clear solution was heated at 80° C. for 12 hrs. Thesolution was concentrated to half the original volume under vacuum, and100 ml of water was added. The solution was reconcentrated, and theprocess was repeated two times (2×100 ml H₂ O). The syrup was placedunder vacuum to remove remaining water and acetic acid. After time, awhite solid formed. The material obtained in quantitative yield was usedwithout further purification; M.P. 128°-130° C.; IR (KBr) 3447, 1613,1570, 1508, 1447, 1316, 1214, 1015, 764 cm⁻¹ ; ¹ H NMR (CDCl₃) δ4.05 (s,3H), 6.91 (d, 1H, J=8.0 Hz), 6.92 (m, 1H), 7.33 (dt, 2H, J=8.0, 1.6 Hz),8.3 (dd, 1H, J=6.3, 1.6 Hz). HRMS Calcd. for C₆ H₇ NO₂ : 125.0477.Found: 125.0474.

Preparation of Intermediate Compound(23)--2-Methoxy-4-nitropyridine-N-oxide

The nitration was carried out using the method of Den Hertog and VanAmmers³. The results obtained from this experiment differ from thosereported. Concentrated H₂ SO₄ (35 ml) was cooled to 0° C., and 15.3 g ofN-oxide (22) (0.12 ml) was cautiously added in portions To this stirredsolution, kept at 0° C., was added the nitrating mixture (conc. H₂ SO₄ ;35 ml: fuming HNO₃ ; 60 ml) dropwise. The ice bath was removed, and themixture was heated to 75° C. for 90 mins. The mixture was recooled to 0°C. and cautiously poured onto 150 g of ice. With vigorous stirring,portions of solid K₂ CO₃ were added until the pH was 7. The liquid wasthen extracted several times with CH₂ Cl₂ (3×200 ml). The aqueous layerwas continuously extracted with CHCl₃. The organic layers were combined,dried over anhydrous Na₂ SO₄ and concentrated to give a yellow solid.The solid was chromatographed on a flash silica column using a gradientsystem of MeOH/CH₂ Cl₂ ; 2:98, 3:97; 4:96; 5:95. A mixture of2-methoxy-4-nitropyridine and 2-methoxy-5-nitropyridine (2.9 g) elutedfirst, followed by 2-methoxy-4-nitropyridine-N-oxide (6.4 g), and then2-methoxy-5-nitropyridine-N-oxide (2.9 g). Compound (23), was obtainedas a yellow solid (30%): M.P. 176°-178° C. (decomp.); (Literature:154.5°-158.5° C., dec.)2; IR (KBr) 3106, 3082, 1601, 1528, 1346, 1296,1231, 1088, 1011, 660 cm⁻¹ ; ¹ H NMR (CDCl₃) δ4.18 (s, 3H), 7.73 (d, 1H,J=2.9 Hz), 7.78 (dd, 1H, J=7.1, 2.9 Hz), 8.35 (d, 1H, J=7.1 Hz). Anal.Calcd. for C₆ H₆ N₂ O₄ : C, 42.36; H, 3.56; N, 16.47. Found: C, 42.42;H, 3.57; N, 16.41.

Preparation of Intermediate Compound(24)--4-Benzylthio-2-methoxypyridine-N-oxide

The pyridine-N-oxide (24) was prepared in similar fashion to thepreparation of compound (19), with the following changes. Once the4-nitro-2-methoxypyridine-N-oxide was added, the reaction mixture wasallowed to warm to room temperature. Stirring was continued for 12 hrs.The precipitated solid that forms was filtered and washed with ice coldTHF. The solid was dried under vacuum and shown to be one spot by TLC(MeOH/CH₂ Cl₂ ; 10:90). The filtrate was concentrated and flashchromatographed on silica with MeOH/CH₂ Cl₂ (gradient: 4:96, 5:95 6:94).An analytically pure tan solid was isolated. The total combined yieldwas 70%. M.P. 131°-133° C.; IR (KBr) 3105, 3038, 3005, 1670, 1610, 1543,1483, 1290, 1211, 1132, 1016, 802 cm⁻¹ ; ¹ H NMR (CDCl₃) δ3.95 (s, 3H),4.19 (s, 2H), 6.64 (d, 1H, J=2.4 Hz), 6.78 (dd, 1H, J=6.9, 2.4 Hz), 7.33(m, 5H), 8.09 (d, 1H, J=6.9 Hz). Anal. Calcd. for C₁₃ H₁₃ NO₂ S: C,63.13; H, 5.30; N, 5.66; S, 12.96. Found: C, 62.88; H, 5.28; N, 5.62; S,12.89.

Preparation of Intermediate Compound(25)--4-Benzylthio-2-methoxypyridine

The starting pyridine-N-oxide (24), (1.85 g) was reduced using themethod to prepare compound (20), except heating of the mixture was notnecessary. The reaction was complete in approximately 90 minutes. Flashsilica chromatography using ether/petroleum ether, 5:95; yielded 1.57 g(90%) of compound (25) as a tan solid. M.P. 35°-36° C.; IR (KBr) 3028,2943, 1589, 1543, 1385, 1307, 1037, 715 cm⁻¹ ; ¹ H NMR (CDCl₃) δ3.98 (s,3H), 4.23 (s, 2H), 6.64 (d, 1H, J=1.6 Hz), 6.84 (dd, 1H, J=5.9, 1.6 Hz),7.35 (m, 5H), 7.98 (d, 1H, J=5.9 Hz). Anal Calcd. for C₁₃ H₁₃ NOS: C,67.50; H, 5.66; N, 6.05; S, 13.86. Found: C, 67.60; H, 5.70; N, 6.10; S,13.80.

Preparation of Compound (26) (Compound16A)--3,4-Dihydro-2,6-dimethyl-4-oxo-5-[4-(6-methoxypyridylthio)]quinazoline

This compound was prepared in 6-7% yield as described for (21) (Compound12A). Tan solid; M.P. 223°-226° C.; IR (KBr) 3445, 1684, 1675, 1669,1452, 1394, 1320, 1038 cm⁻¹ ; ¹ H NMR (DMSO-d₆) δ2.28 (s, 3H), 2.35 (s,3H), 3.70 (s, 3H), 6.05 (s, 1H), 6.49 (dd, 1H, J=4.1, 2.9 Hz), 7.60 (d,1H, J=8.5 Hz), 7.78 (d, 1H, J=8.4 Hz), 7.85 (d, 1H, J=5.4 Hz), 12.10 (s,1H). HRMS calcd. for C₁₆ H₁₅ N₃ O₂ S: 313.0885. Found: 313.0882.

Example 5

Preparation of Compounds 17A and 18A

Compounds 17A and 18A were prepared according to the following reactionscheme: ##STR69## Preparation of Intermediate Compound(27)--2-Trifluoromethylpyridine-N-oxide

Using the procedure to prepare intermediate (22),2-trifluoromethylpyridine-N-oxide was synthesized in 72% yield startingfrom 2-trifluoromethylpyridine. (Yellow oil); IR (neat) 3125, 3085,1721, 1615, 1439, 1329, 1269, 1115, 1071, 1044, 852, 771, 662 cm⁻¹, ¹ HNMR (CDCl₃) δ7.38 (t, 1H, J=7.9 Hz), 7.48 (dt, 1H, J=7.0, 2.1 Hz), 7.71(dd, 1H, J=7.9, 2.1 Hz), 8.35 (d, 1H, J=6.5 Hz). Anal. Calcd. for C₆ H₄F₃ NO.0.5 H₂ O: C, 41.87; H, 2.93; F, 33.12; N, 8.14. Found: C, 41.84;H, 2.81; F, 33.19; N, 8.26.

Preparation of Intermediate Compound(28)--4-Nitro-6-trifluoromethylpyridine-N-oxide

The nitration of pyridine-N-oxide (27) was carried out using the samemethod to prepare compound (23), with the following changes. Thereaction mixture was heated at 125°-130° C. for 31/2 hrs. Duringwork-up, no continuous extraction of the aqueous layer was necessary.The crude solid was purified employing flash column chromatography onsilica using ethyl acetate/hexane; 20:80. The product was isolated as ayellow solid (M.P. 112°-114° C.) in 38% yield. IR (KBr) 3416, 3125,1620, 1591, 1537, 1449, 1354, 1306, 1281, 1165, 1130, 916, 693 cm⁻¹ ; ¹H NMR (CDCl₃) δ8.28 (dd, 1H, J=7.2, 3.1 Hz), 8.36 (d, 1H, J=7.2 Hz),8.52 (d, 1H, J=3.1 Hz). Anal. Calcd. for C₆ H₃ F₃ N₂ O₃ : C, 34.63; H,1.45; F, 27.39; N, 13.46. Found: C, 34.86; H, 1.35; F, 27.16; N, 13.66.

Preparation of Intermediate Compound(29)--4-Amino-6-trifluoromethylpyridine

In a Parr hydrogenation bottle, 8.32 g of nitropyridine-N-oxide (28)(0.04 mol) was dissolved in 275 ml of 95% ethanol. The bottle wasflushed with argon, and 0.83 g of 10% palladium on activated carbon wasadded. The bottle was shaken under 35 psi of hydrogen for 45 min on aParr hydrogenator. At this time, the catalyst was filtered off through acelite pad. The ethanolic filtrate was concentrated under vacuo, and theoil was dissolved in 50 ml of dichloromethane. This solution wasfiltered through a small pad of silica gel to remove traces of catalystand carbon. The filtrate was concentrated, and traces of solvent wereremoved under vacuo. The oil slowly crystallized to give 5.77 g (89%yield) of an analytically pure light orange solid. M.P. 56°-58° C.; IR(KBr) 3501, 3335, 3175, 1657, 1611, 1472, 1373, 1300, 1169, 1117, 993,850 cm⁻¹ ; ¹ H NMR δ4.40 (bs, 2H), 6.64 (dd, 1H, J=5.6, 2.3 Hz), 6.89(d, 1H, J=2.3 Hz), 8.30 (d, 1H, J=5.6 Hz). Anal. Calcd. for C₆ H₅ F₃ N₂: C, 44.45; H, 3.11; F, 35.16; N, 17.28. Found: C, 44.56; H, 2.95; F,35.14; N, 17.28. HRMS calcd. for C₆ H₅ F₃ N₂ : 162.0405. Found:162.0402.

Preparation of Intermediate Compound(30)--Ethyl-4-(6-trifluoromethylpyridyl)xanthate

A solution of 4.86 g of Amine (29) (0.03 mol) in 30 ml of concentratedH₂ SO₄ was cooled to 0° C. An aqueous solution (30 ml H₂ O) of 2.69 gNaNO₂ (39.0 mmol) was cooled to 0° C. and added dropwise over a periodof 15 min. Stirring of the broken mixture was continued at 0° C. for 5additional minutes. At this time, an ice cold solution of 8.17 g ofpotassium ethyl xanthate (51.0 mmol) in 30 ml of H₂ O was addeddropwise, maintaining the reaction temperature, between 0°-5° C. Themixture was warmed to room temperature, and dichloromethane (125 ml) wasadded. The aqueous layer was neutralized to pH7 with solid Na₂ CO₃. Theorganic layer was separated, and the aqueous layer was extracted withethyl acetate (3×50 ml). The organic layers were combined, dried overanhydrous Na.sub. 2 SO₄, and concentrated. The residue was subjected toflash column chromatography using silica gel and a gradient solventsystem of ethyl acetate/hexane (2:98, 2.5:97.5, 3:97). Compound (30) wasisolated as a yellow oil in 36% yield and was used without furtherpurification. IR (neat) 3061, 2988, 2901, 1738, 1584, 1555, 1406, 1323,1252, 1184, 1146, 1038, 845, 720 cm⁻¹ ; ¹ H NMR (CDCl₃) δ1.38 (t, 3H,J=7.1 Hz), 4.66 (q, 2H, J=7.1 Hz), 7.60 (dd, 1H, J=5.0, 1.3 Hz), 7.83(d, 1H, J=1.0 Hz), 8.77 (d, 1H, J=5.0 Hz). HRMS calcd. for C₉ H₈ F₃ NOS₂(M+1): 268.0077. Found (M+1): 268.0065.

Preparation of Compound 31 (Compound17A)--3,4-Dihydro-2,6-dimethyl-4-oxo-5-[4-(6-trifluoromethylpyridylthio)]-quinazoline

To a solution of 0.67 gm of xanthate (30) (2.5 mmol) in 3 ml MeOH wasadded 2.5 ml 1N KOH in methanol, and the mixture was stirred for 11/2hr. The mixture was evaporated to dryness, and to the residue was added10 ml of anhydrous N,N-Dimethylacetamide, 0.25 g quinazoline (6) (10.0mmol), 0.1 g copper (I) bromide, and 0.1 g copper (I) oxide. The mixturewas heated at 90° C. for 6 hrs. and then the solvent was evaporated. Thesolid was treated with 50 ml of H₂ S/MeOH solution (20 g/l) for 1 hr.The mixture was filtered, and the filtrate was evaporated to dryness.The solid was purified via flash chromatography on silica using MeOH/CH₂Cl₂ (5:95) to yield 65 mg (18.5% theory) of a yellow solid: M.P.240°-245° C.; IR (KBr) 3440, 3190, 3057, 2950, 1675, 1630, 1595, 1321,1140, 720 cm⁻¹ ; ¹ H NMR (DMSO-d6) δ2.28 (s, 3H), 2.42 (s, 3H), 6.97 (d,1H, J=5.2 Hz), 7.46 (d, 1H, J=1.1 Hz), 7.67 (d, 1H, J=8.4 Hz), 7.84 (d,1H, J=8.4 Hz), 8.37 (d, 1H, J=5.2 Hz), 12.05 (bs, 1H). HRMS. Calcd. forC₁₆ H₁₂ F₃ N₃ OS: 351.0656. Found: 351.0653.

Preparation of Compound 32 (Compound18A)--2-Amino-3,4-dihydro-6-methyl-4-oxo-5-[4-(6-trifluoromethylpyridylthio)]-quinazoline

This compound was prepared in 22% yield as described above. Tan solid;M.P. 247°-249° C.; IR (KBr) 3421, 2056, 1650, 1625, 1485, 1419, 1328,1146, 815, 724 cm⁻¹ ; ¹ H NMR (DMSO-d₆) δ2.30 (s, 3H), 6.50 (bs, 2H),6.97 (dd, 1H, J=4.1, 1.2 Hz), 7.30 (d, 1H, J=8.4 Hz), 7.39 (d, 1H, J=1.0Hz), 7.62 (d, 1H, J=8.6 Hz), 8.36 (d, 1H, J=5.2 Hz), 12.10 (bs, 1H).HRMS calcd. for C₁₅ H₁₁ F₃ N₄ OS (M+1): 353.0677. Found (M+1): 353.0684.

Example 6

Preparation of Compound 26A

Compound 26A was prepared according to the following reaction scheme:##STR70## Preparation of Intermediate Compound(33)--6-Dimethylamino-4-nitropyridine-N-oxide

To a solution of 5.0 g (23 mmol) of 2-Bromo-4-nitropyridine-N-oxide⁴dissolved in 75 ml of tetrahydrofuran, was added 1.1 g (24 mmol)dimethylamine. The mixture was stirred for 3 hrs, followed by filtrationto remove the dimethylamine-hydrobromide salt. The filtrate wasevaporated to dryness, and the crude solid was purified by flash columnchromatography on silica using methanol/dichloromethane; 4:96. Theproduct was isolated as an orange solid (M.P. 128°-130° C.) in 83%yield. ¹ NMR (CDCl₃) δ3.14 (s, 6H), 7.67 (m, 2H), 8.24 (d, 1H, J=7.1Hz). Anal. Calcd. for C₇ H₉ N₃ O₃ : C, 45.60; H, 4.95; N, 22.94. Found:C, 46.00; H, 5.00; N, 22.96.

Preparation of Intermediate Compound(34)--6-Dimethylamino-4-(4-methoxybenzylthio)-pyridine-N-oxide

To a solution of 1.1 g (7.1 mmol) of 4-Methoxy-α-toluenethiol dissolvedin 75 ml anhydrous DMF, was added 0.28 g (7.0 mmol; 60 wt. % dispersionin mineral oil). After stirring for 1 hr, a solution of 1.2 g (6.55mmol) of pyridine-N-oxide 33 in 25 ml anhydrous DMF was added dropwise.The reaction mixture was stirred for 2 hrs and was then poured into 200ml. H₂ O. The aqueous solution was extracted with 500 ml diethyl ether,separated, and dried over anhydrous MgSO₄. The ether was evaporated togive compound 34 as a tan solid in 63% yield. ¹ H NMR (CDCl₃) δ3.06 (s,6H), 3.83 (s, 3H), 4.16 (s, 2H), 6.60 (d, 1H, J=2.5 Hz), 6.70 (d, 1H,J=7.0 Hz), 6.90 (d, 2H, J= 8.7 Hz), 7.30 (d, 2H, J=8.7 Hz), 8.0 (d, 1H,J=7.0 Hz).

Preparation of Intermediate Compound(35)--6-Dimethylamino-4-(4-methoxybenzylthio)-pyridine

The starting pyridine-N-oxide 34, (0.60 g; 2.07 mmol) was reduced usingthe method to prepare compound 25. Upon completion of the reaction, themixture was poured into 200 ml H₂ O, and the pH was adjusted to 7. Theaqueous solution was extracted with ethyl acetate (500 ml), and theorganic phase was dried over anhydrous Na₂ SO₄, filtered andconcentrated. No chromatography was necessary, and the product, 35, wasisolated as a yellow solid in 88% yield. ¹ H NMR (CDCl₃) δ3.08 (s, 3H,3.82 (s, 3H), 4.17 (s, 2H, 6.34 (d, 2H, J=1.3 Hz), 6.48 (d, 1H, J=5.5Hz), 6.87 (d, 2H, J=8.7 Hz), 7.33 (d, 2H, J=8.7 Hz), 8.0 (d, 1H, J=5.5Hz).

Preparation of Intermediate Compound(36)--6-Dimethylamino-4-mercaptopyridine

A formic acid (10 ml) solution of pyridine 35 (0.40 g; 1.46 mmol) wascooled to 0° C. To this solution was added 1.2 g of Hg(OAc)₂ dissolvedin 3 ml H₂ O. The ice bath was removed, and the reaction mixture wasallowed to stir for 12 hours. At this time, the pH was adjusted by theaddition of aqueous ammonia. A grey precipitate formed which wasfiltered, washed with an excess of H₂ O and air dried. The solid wasthen taken up in a saturated H₂ S/methanol solution. A black solid (HgS)formed and was filtered off. The filtrate was evaporated to dryness toyield a yellow solid (89%) which was used without further purification.¹ H NMR (CDCl₃) δ3.12 (s, 3H), 3.5 (bs, 1H), 6.5 (d, 1H, J=5.3 Hz), 6.57(d, 1H, J=3.6 Hz), 7.69 (d, 1H, J=5.5 Hz).

Preparation of Compound 37 (Compound26A)--3,4-Dihydro-2,6-dimethyl-4-oxo-5-[4-(6-dimethylaminopyridylthio)]-quinazoline

This compound was prepared from intermediates 6 and 36 using the exactprocedure to generate compound 9 (8A). The crude product was purified byflash column chromatography on silica gel using MeOH/CH₂ Cl₂ (8:92) togive a tan solid in 21% yield. ¹ NMR (DMSO-d₆) δ1.97 (s, 3H), 2.08 (s,3H), 2.56 (s, 3H), 5.55 (d, 1H, J=5.4 Hz), 5.82 (d, 1H, J=1.2 Hz), 7.27(d, 1H, J=8.4 Hz), 7.44 (d, 1H, J=5.2 Hz), 7.45 (d, 2H, J=8.4 Hz), 12.75(bs, 1H). Anal. Calcd. for C₁₇ H₁₈ N₄ OS.0.5H₂ O: C, 60.82; H, 5.66; N,16.69; S, 9.54. Found: C, 61.01; H, 5.63; N, 16.55; S, 9.42.

Example 7

Preparation of Compounds 27A and 28A

Compounds 27A and 28A were prepared according to the following reactionscheme: ##STR71## Preparation of Intermediate Compound(38)--Methyl-4-[3,4-dihydro-2,6-dimethyl-4-oxo-5-quinazolinyl)thio]-benzoate

This compound was prepared from intermediate 6 andmethyl-4-mercaptobenzoate [P. R. Marsham et al., J. Med. Chem. 34 2209(1991); and E. Campaigne, et al., J. Org. Chem. 27 2835 (1962)] usingthe procedure to synthesize compound 9 (8A). After heating the mixtureat 90° C. for 16 hrs, the DMA was removed under vacuum, and the solidresidue was suspended in methanol. To this stirred suspension, a streamof gaseous H₂ S was bubbled slowly in for approximately 5 min. A darksolid (CuS) formed which was removed by filtration. The methanolicfiltrate was concentrated, and the product was purified by flash columnchromatography on silica with methanol/dichloromethane; 5:95; to give atan solid in 85% yield. ¹ H NMR (DMSO-d₆) δ2.26 (s, 3H), 2.43 (s, 3H),3.75 (s, 3H), 6.94 (d, 2H, J=8.4 Hz), 7.22 (d, 1H, J=8.5 Hz), 7.52 (d,1H, J=8.5 Hz), 7.71 (d, 2H, J=8.4 Hz), 11.7 (bs, 1H). HRMS calcd. forC₁₈ H₁₆ N₂ O₃ S: 340.0898. Found: 340.0882.

Preparation of Compound (39) (Compound28A)--4-[(3,4-Dihydro-2,6-dimethyl-4-oxo-5-quinazolinyl)thio]-benzoicacid

An ethanolic solution (5 ml) consisting of 0.186 g (0.55 mmol) of methylester 38 and 0.5 ml of aqueous 1N NaOH was heated at 50° C. for 4 hrs.At this time, the solution was evaporated to dryness, and the sodiumsalt was dissolved in 3 ml H₂ O. This solution was carefully acidifiedto pH 4 with concentrated HCl. The free acid which precipitated wasfiltered and washed with 5 ml of cold H₂ O. The solid was dried in adesicator over CaSO₄ to yield 0.15 g (84%) of acid 39 (28A) as a beigesolid. ¹ H NMR (DMSO-d₆) δ2.29 (s, 3H), 2.45 (s, 3H), 7.0 (d, 2H, J=8.5Hz), 7.44 (d, 1H, J=8.6 Hz), 7.71 (d, 2H, J=8.4 Hz), 7.74 (d, 1H, J=8.3Hz). HRMS calcd. for C₁₇ H₁₄ N₂ O₃ S: 326.0742. Found: 326.0725.

Preparation of Intermediate Compound(40)--Diethyl-N-[4-((3,4-dihydro-2,6-dimethyl-4-oxo-5-quinazolinyl)thio)benzoyl]-L-glutamate

Benzoic acid 39 (60.0 mg; 18.4 mmol) and (L)-glutamic acid diethylester.HCl (0.144 g; 0.6 mmol) were dissolved in 5 ml of anhydrous DMFand cooled to 0° C. To the stirred solution was added diphenylphosphorylazide (0.15 ml; 0.7 mmol). After 15 min, 0.2 ml (1.4 mmol) oftriethylamine was added, and the reaction mixture was allowed to stirfor 12 hrs at room temperature. The solvent was then removed undervacuum, and the remaining solid was taken up in 5 ml H₂ O. The pH wascarefully adjusted to 6 with concentrated HCl, and the aqueous solutionwas extracted with CDCl₃ (3×10 ml). The organic layers were combined,dried over MgSO₄, filtered and evaporated to dryness. The product waspurified by flash chromatography on silica usingmethanol/dichloromethane; 10:90. A tan solid (78.0 mg; 82%) wasisolated. ¹ H NMR (DMSO-d₆) δ1.11 (m, 6H), 1.61 (m, 2H), 1.79 (m, 2H),2.26 (s, 3H), 2.37 (s, 3H), 3.26 (m, 1H), 4.05 (m, 4H), 6.96 (d, 2H,J=8.4 Hz), 7.55 (d, 1H, J=8.5 Hz), 7.64 (d, 2H, J=8.4 Hz), 7.71 (d, 1H,J=8.5 Hz), 8.60 (d, 1H, J=5.3 Hz), 12.10 (bs, 1H). HRMS calcd. for C₂₆H₂₉ N₃ O₆ S (M+1): 512.1843. Found (M+1): 512.1855.

Preparation of Compound (41) (Compound27A)--N-[4-((3,4-Dihydro-2,6-dimethyl-4-oxo-5-quinazolinyl)thio)benzoyl]-L-glutamicacid

Diethyl ester 40 (78.0 mg; 0.15 mmol) was dissolved in 5 ml of ethanol,and to this solution was added 0.5 ml of an aqueous 1N NaOH solution.The reaction mixture was stirred at 50° C. for 3 hrs, where upondisappearance of starting by TLC the solution was evaporated to dryness.The disodium salt was further dissolved in 2 ml of H₂ O and acidified topH 4 with concentrated HCl. The solid was filtered upon precipitationand washed with 5 ml of cold H₂ O. The final product was dried undervacuum over CaSO₄ yielding 50 mg (72%) of an off white solid. ¹ H NMR(DMSO-d₆) δ1.95 (m, 2H), 2.05 (m, 2H), 2.26 (s, 3H), 2.46 (s, 3H), 4.40(m, 1H), 6.93 (d, 2H, J=8.4 Hz), 7.55 (d, 1H, J=8.4 Hz), 7.65 (d, 2H,J=8.5 Hz), 7.71 (d, 1H, J=8.4 Hz), 8.40 (bd, 1H, J=5.4 Hz), 12.00 (bs,1H). Anal. Calcd. for C₂₂ H₂₁ N₃ O₆ S.2HCl: C, 50.05; H, 4.36; N, 7.96;S, 6.06. Found: C, 50.38; H, 4.69; N, 7.60; S, 5.77.

Example 8

Preparation of Compounds 3A and 5A

Compounds 3A and 5A were prepared according to the following reactionscheme: ##STR72## Preparation of Intermediate Compound(42)--3,4-Dihydro-2,5-dimethyl-4-oxo-quinazolinone.

This compound was prepared via its corresponding benzoxazinone from6-methylanthranilic acid using the procedure to prepare quinazolinone(6). The solid was recrystallized from ethanol. (M.P. 258°-259° C.). ¹ HNMR (CDCl₃) δ2.53 (s, 3H), 2.89 (s, 3H), 7.20 (d, 1H, J=7.3 Hz), 7.50(d, 1H, J=8.0 Hz), 7.59 (dd, 1H, J=8.1, 7.3 Hz), 11.52 (bs, 1H). Anal.Calcd. for C₁₀ H₁₀ N₂ O: C, 68.95; H, 5.79; N, 16.08. Found: C, 69.03;H, 5.82; N, 16.03.

Preparation of Intermediate Compound(43)--2,5-Dimethyl-3-[2'-(trimethylsilyl)ethoxymethyl]-quinazolin-4-one.

To 70 ml of dry DMF was added 2.175 g (12.5 mmol) of quinazolinone (42).The mixture was cooled to 0° C., and 0.55 g of NaH (13.75 mmol; 60% oildispersion) was added portionwise with stirring. The green coloredmixture was allowed to warm to room temperature, and stirring wascontinued until gas (H₂) evolution ceased. At this time, the solutionwas recooled to 0° C., and2-(Trimethylsilyl)ethoxymethylchloride(SEM-Cl) (2.45 ml; 13.75 mmol) wasadded dropwise. A cloudy precipitate (NaCl) began to form. After all theSEM-Cl was added, the ice bath was removed, and the reaction mixture wasstirred at room temperature for 12 hrs. The mixture was poured into H₂ O(300 ml) and extracted with hexanes (3×150 ml). The organic layers werecombined and dried over anhydrous MgSO₄. Upon filtration andconcentration, a white powder began to form. The solid was removed byfiltration and was shown to be the starting material, (42), by TLC and ¹H NMR. The filtrate was concentrated to give a pale yellow oil which waspassed through a flash silica gel column using diethyl ether/petroleumether; 1:1, yielding 3.0 g (79%) of product (43) as an oil. IR (neat)2980, 1675, 1600, 1572, 1460, 1380, 1287, 1248, 1075, 858, 835 cm⁻¹ ; ¹H NMR (CDCl₃) δ0.00 (s, 9H), 0.95 (dd, 2H, J=8.2, 7.2 Hz), 2.66 (s, 3H),2.84 (s, 3H), 3.68 (dd, 2H, J=8.2, 7.1 Hz), 5.52 (s, 2H), 7.18 (d, 1H,J=6.8 Hz), 7.43 (dd, 1H, J=8.0, 0.3 Hz), 7.55 (dd, 1H, J=8.0, 7.5 Hz).

Preparation of Intermediate Compound(44)--5-Bromomethyl-2-methyl-3-[2'-(trimethylsilyl)ethoxymethyl]-quinazolin-4-one.

The SEM protected quinazolinone (43) (2.28 g, 7.5 mmol) was dissolved in30 ml CCl₄. To the solution was added 1.47 g (8.23 mmol) ofN-bromosuccinimide. The pale yellow solution was heated to a gentlereflux until almost completely homogeneous. At this time, the benzylicbromination reaction was initiated with a 200 watt lamp. The reactionbegan to reflux more vigorously and turned a deep orange color. Afterapproximately 15 min, the color faded and succinimide precipitated. Thereaction was cooled, filtered and washed with 25 ml CCl₄. The filtratewas washed with minimal H₂ O (˜5 ml), separated, dried over MgSO₄,refiltered and concentrated leaving a solid residue which was furtherpurified by flash column chromatography on silica using a gradientsystem of diethyl ether/petroleum ether; 15/85; 20/80; 25/75; 30/70;35/65. The pure bromide (1.25 g) was isolated as a white sold in 43%yield (54% based on recovered (43): M.P. 78°-80° C. IR (KBr) 3085, 2980,1675, 1608, 1382, 1340, 1293, 1248, 1075, 860, 830, 710 cm⁻¹ ; ¹ H NMR(CDCl₃) δ0.00 (s, 9H), 0.93 (dd, 2H, J=8.3, 7.1 Hz), 2.66 (s, 3H), 3.68(dd, 2H, J=8.4, 7.1 Hz), 5.24 (s, 2H), 5.55 (s, 2H), 7.38 (dd, 1H,J=7.2, 1.5 Hz), 7.55 (dd, 1H, J=7.5, 1.5 Hz), 7.61 (dd, 1H, J=7.5, 7.2Hz). Anal. Calcd. for C₁₆ H₂₃ BrN₂ O₂ Si: C, 50.12; H, 6.04; Br, 20.84;N, 7.30. Found: C, 50.35; H, 6.06; Br, 21.01; N, 7.32.

Preparation of Intermediate Compound(45)--5-Chloro-N-[2'-methyl-3'-(2"-(trimethylsilyl)ethoxymethyl)-4'-oxo-5'-quinazolyl)methyl]indole.

In 6.5 ml anhydrous DMF, 0.417 g (2.75 mmol) of 5-chloroindole wasdissolved. The stirred solution was cooled to 0° C., and 0.11 g (2.75mmol, 60% oil dispersion) of NaH was added portionwise. Once the anionwas formed (˜30 min.), 0.958 g (2.5 mmol) of Bromomethylquinazoline(44), dissolved in 0.5 ml anhydrous DMF was syringed in. The reactionwas complete upon disappearance of starting materials by TLC(40%ether/petroleum ether). Ice was added to quench excess anion, followedby 20 ml H₂ O. This was then extracted with diethyl ether (3×50 ml), andthe organic layers were combined and dried over anhydrous MgSO₄.Filtration and evaporation gave a residue which was purified by flashcolumn chromatography on silica using diethyl ether/petroleum ether:40:60. A white crystalline solid was isolated (0.927 g; 82%; M.P.98°-99° C.). IR (KBr) 3095, 2980, 1715, 1595, 1565, 1440, 1345, 1280,1245, 1175, 1060, 932, 834, 795, 755, 720, 612 cm⁻¹ ; ¹ H NMR (CDCl₃)δ0.03 (s, 9H), 1.00 (m, 2H), 2.71 (s, 3H), 3.74 (m, 2H), 5.55 (s, 2H),6.04 (s, 2H), 6.25 (dd, 1H, J=7.4, 1.3 Hz), 6.54 (dd, 1H, J=3.1, 0.3Hz), 7.08 (m, 2H), 7.18 (d, 1H, J=3.1 Hz), 7.45 (m, 2H), 7.63 (dd, 1H,J=1.6, 1.0 Hz). Anal. Calcd. for C₂₄ H₂₈ ClN₃ O₂ Si: C, 63.48; H, 6.21;Cl, 7.80; N, 9.25. Found: C, 63.41; H, 6.13; Cl, 7.91; N, 9.19.

Preparation of Compound (46) (Compound3A)--5-Chloro-N-[(3,4-dihydro-2-methyl-4-oxo-5-quinazolyl)methyl]indole.

The SEM protected quinazoline (45) (0.75 g; 1.65 mmol) was dissolved in1.5 ml THF. To this solution was added 6.0 ml of a 1.0M THF solution oftetrabutylammonium fluoride. With stirring, the mixture was heated to50° C. for 7 hrs. The solution was cooled to room temperature, and 20 mlof H₂ O was added. This was then extracted with a large excess (200 ml)of ethyl acetate. The organic layer was separated and dried overanhydrous MgSO₄, filtered and concentrated leaving a solid residue whichwas recrystallized from ethyl acetate yielding product (46) (Compound3A) in 46%. M.P. 251°-252° C.; ¹ H NMR (DMSO-d₆) δ2.34 (s, 3H), 6.08 (s,2H), 6.14 (dd, 1H, J=7.3, 1.0 Hz), 6.53 (dd, 1H, J=3.0, 0.5 Hz), 7.05(dd, 1H, J=8.7, 2.1 Hz), 7.34 (d, 1H, J=8.8 Hz), 7.46 (m, 2H), 7.56 (d,1H, J=3.1 Hz), 7.64 (d, 1H, J=2.1 Hz), 12.30 (bs, 1H). Anal. Calcd. forC₁₈ H₁₄ ClN₃ O.0.1 EtOAc: C, 66.45; H, 4.49; Cl, 10.66; N, 12.63. Found:C, 66.68; H, 4.62; Cl, 10.95; N, 12.27. HRMS Calcd. for C₁₈ H₁₄ ClN₃ O:323.0825. Found: 323.0813.

Preparation of Intermediate Compound(47)--5-Formyl-2-methyl-3-[2'-(trimethylsilyl)ethoxymethyl]-quinazolin-4-one.

To a solution of NaOEt in ethanol, prepared by dissolving 34.5 mg (1.5mmol) of sodium metal in 1.5 ml of absolute ethanol, was added 0.14 ml(1.56 mmol) of 2-nitropropane. 0.575 g (1.5 mmol) ofBromomethylquinazoline (44) was added, and the reaction was stirred andheated at 40° C. for 12 hrs. At this time, 10 ml of H₂ O was added, andthe mixture was extracted with diethyl ether (2×50 ml). The organiclayers were separated, dried over anhydrous Na₂ SO₄, filtered andconcentrated. The residue was purified by flash column chromatographywith 70% ether/pet. ether yielding 0.346 g (73%) of aldehyde (47) as awhite solid. ¹ H NMR (CDCl₃) δ0.00 (s, 9H), 0.96 (m, 2H), 2.73 (s, 3H),3.75 (m, 2H), 5.58 (s, 2H), 7.82 (m, 2H), 7.87 (m, 1H), 11.17 (s, 1H).

Preparation of Intermediate Compound(48)--5-(α-Hydroxytolyl)-2-methyl-3-[2'-(trimethylsily)ethoxymethyl]-quinazolin-4-one.

Aldehyde (47) (0.72 g; 2.26 mmol) was dissolved in 9.0 ml of anhydrousTHF under an argon atmosphere. The stirred solution was cooled to -78°C., and phenylmagnesium bromide (0.83 ml; 3.0M in diethyl ether) wasintroduced dropwise. The reaction mixture was allowed to warm to roomtemperature, and stirring was continued for 1 hr. To quench thereaction, 10 ml of sat. aq. NH₄ Cl was added. The mixture was thenextracted with diethyl ether (3×50 ml), separated, combined, dried overanhydrous MgSO₄, filtered and concentrated. Purification of the residueby flash column chromatography on silica with ether/pet. ether; 60:40;supplied 0.621 g of the benzylic alcohol (48), as a colorless oil in 74%yield. IR (neat) 3380, 3070, 3035, 2960, 2900, 1660, 1600, 1540, 1445,1245, 1135, 1075, 915, 830, 695 cm⁻¹ ; ¹ H NMR (CDCl₃) δ0.00 (s, 9H),0.88 (m, 2H), 2.68 (s, 3H), 3.54 (m, 2H), 5.50 (s, 2H), 5.77 (d, 1H,J=8.3 Hz), 6.36 (d, 1H, J=8.0 Hz), 7.25 (m, 4H), 7.30 (m, 2H), 7.60 (dd,1H J=8.2, 1.5 Hz), 7.67 (dd, 1H, J=7.7, 7.2 Hz). Anal. Calcd. for C₂₂H₂₈ N₂ O₃ Si: C, 66.63; H, 7.11; N, 7.06. Found: C, 66.66; H, 6.97; N,7.00.

Preparation of Compound (49) (Compound5A)--3,4-Dihydro-2-methyl-4-oxo-5-(α-hydroxytolyl)-quinazoline.

Following the same procedure to prepare compound (46) (Compound 3A), theSEM-quinazoline (48) was deprotected using 3.0 eq. of tetrabutylammoniumfluoride at 50° C. for 4 hrs. Quinazoline (49) (Compound 5A) wasisolated as a white solid in 38% yield after purification by flashchromatography on silica using methanol/dichlormethane; 5:95. ¹ H NMR(DMSO-d6) δ2.29 (s, 3H), 5.93 (d, 1H, J=5.1 Hz), 7.15 (m, 1H), 7.21 (m,3H), 7.27 (m, 2H), 7.45 (dd, 1H, J=5.6, 3.9 Hz), 7.73 (m, 2H), 12.06(bs, 1H).

Example 9

Preparation of Compounds 4A and 6A

Compounds 4A and 5A were prepared according to the following reactionscheme: ##STR73## Preparation of Intermediate Compound(50)--5-Benzoyl-2-methyl-3-[2'-(trimethylsilyl)ethoxymethyl]-quinazolin-4-one.

Benzylic alcohol (48) (0.569 g; 1.43 mmol) was taken up in 18 ml of dryCH₂ Cl₂ and stirred under an inert atmosphere. Activated MnO₂ (1.43 g)was added, and the progress of the reaction was followed by TLC(ether/pet. ether; 70/30). Upon disappearance of starting material, theblack mixture was filtered through a pad of celite, and the pad waswashed thoroughly with CH₂ Cl₂ (100 ml). The filtrate was dried overanhydrous Na₂ SO₄, filtered and concentrated, leaving 0.48 g (85%) as awhite solid that was analytically pure. M.P 124°-125° C.; IR (KBr) 3010,2955, 2900, 1660, 1560, 1440, 1345, 1292, 1245, 1178, 1060, 920, 825,685 cm⁻¹ ; ¹ H NMR (CDCl₃) δ-0.10 (s, 9H), 0.82 (m, 2H), 2.70 (s, 3H),3.48 (m, 2H), 5.42 (s, 2H), 7.31 (dd, 1H, J=7.1, 1.3 Hz), 7.39 (m, 2H),7.51 (m, 1H), 7.72 (m, 1H), 7.74 (m, 2H), 7.80 (dd, 1H, J=8.2, 7.1 Hz).Anal. Calcd. for C₂₂ H₂₆ N₂ O₃ Si: C, 66.97; H, 6.64; N, 7.10. Found: C,66.76; H, 6.52; N, 6.95.

Preparation of Compound (51) (Compound4A)--5-Benzoyl-3,4-dihydro-2-methyl-4-oxo-quinazoline.

The protected quinazoline (50) (0.255 g; 0.64 mmol) was added to 7.0 mlof 1:1 THF: 2N HCl. The mixture became homogeneous when heated Justunder reflux. After 3 hrs, a white precipitate formed. The mixture wascooled to room temperature, and 10 ml of cold H₂ O was added. Withvigorous stirring, an excess of sat. aq. NaHCO₃ was added. The solid wasfiltered off and washed thoroughly with cold H₂ O (2×10 ml). The solidwas dried under vacuum over activated silica gel desiccant. Theanalytically pure white solid (0.14 g) was isolated in 83% yield. M.P.288°-289° C.; IR (KBr) 3175, 3030, 2880, 1660, 1635, 1325, 1265, 880,825, 780, 725 cm⁻¹ ; ¹ H NMR (DMSO-d₆) δ2.35 (s, 3H), 7.30 (dd, 1H,J=7.3, 1.0 Hz), 7.44 (m, 2H), 7.58 (m, 3H), 7.71 (dd, 1H, J=8.3, 1.0Hz), 7.85 (dd, 1H, J=8.1, 7.3 Hz), 12.22 (bs, 1H). Anal. Calcd. for C₁₆H₁₂ N₂ O₂ : C, 72.71; H, 4.57; N, 10.60. Found: C, 72.61; H, 4.70; N,10.39.

Preparation of Compound (52) (Compound6A)--3,4-Dihydro-5-(α-diphenyl-hydroxymethyl)-2-methyl-4-oxo-quinazoline.

Under an argon atmosphere, 79.3 mg (0.3 mmol) of ketone (51) (Compound4A) was suspended in 5.0 ml of anhydrous THF. The stirred suspension wascooled to 0° C., and 0.375 ml of phenyllithium (2.0M in70:30/cyclohexane:ether) was syringed into the reaction vessel dropwise.Upon addition of the first equivalent of reagent, the starting substratesolubilized. When the addition was completed, the ice bath was removed,and the reaction was allowed to stir for 1 hr. The solution was quenchedwith ice-H₂ O (˜2.0 ml) and poured into 50 ml of CH₂ Cl₂. The aqueouslayer was extracted two more times with CH₂ Cl₂ (50 ml), the organiclayers were combined and dried over anhydrous MgSO₄. The drying agentwas filtered off, and the filtrate was concentrated and purified byflash column chromatography on silica with CH₃ OH:CH₂ Cl₂ /4:96. Theproduct was isolated as a white solid (57 mg; 58%). ¹ H NMR (DMSO-d₆)δ2.33 (s, 3H), 6.55 (dd, 1H, J=5.4, 3.6 Hz), 7.07 (m, 4H), 7.23 (m, 6H),7.59 (m, 2H), 8.73 (s, 1H), 12.46 (bs, 1H). HRMS Calcd. for C₂₂ H₁₈ N₂O₂ : 342.1368. Found: 342.1366.

BIOCHEMICAL AND BIOLOGICAL EVALUATION

Determination of Inhibition Constants Against5,10-Methylenetetrahydrofolate for the Enzyme Thymidylate Synthase

Thymidylate synthase activity was measured using a modification of thetritium release method of Lomax and Greenberg [M. I. S. Lomax and G. R.Greenberg, J. Biol. Chem. 242 109 (1967)]. Inhibition constants, K_(i),slope and K_(i), intercept [W. W. Cleland, Biochim. Biophys. Acta 67 173(1963)], were determined against the cofactor (6R,6S)-5,10-methylene-tetrahydrofolate which was generated in situ byreaction of tetrahydrofolate with formaldehyde [R. G. Kallen and W. P.Jencks, J. Biol. Chem. 241 5851 (1966)]. The cofactor was present as thevariable substrate under conditions of saturating radiolabelled2'-deoxyuridine 5-monophosphate (dUMP). Assays in a total volume of 0.1mL contained 50 mM Tris @ pH 7.6, 10 mM DTT (dithiothreitol), 1 mM EDTA(ethylenediaminetetraacetic acid), 25 mM MgCl₂, 15 mM formaldehyde, ±1%DMSO (depending on the solubility of the compound), 25 μM [5-³ H] dUMP(specific activity 2×10⁸ cpm/μmol), tetrahydrofolate (eightconcentrations ranging from 5 μM to 300 μM) and enzyme (=30 ng for E.coli TS and =60 ng for human TS). Assays of human TS also contained 1-5μg/mL bovine serum albumin to stabilize the protein. Reactions wereinitiated by the addition of enzyme and were carried out for 5 minutesat 24° C., and then quenched by the addition of charcoal (15 mg in 0.1mL H₂ O). The quenched samples were centrifuged at 10,000 rpm for 12-15min at 40° C. to remove unreacted dUMP which had bound to the charcoal,and 0.1 mL of the supernatant was counted by liquid scintillation in thepresence of 5 mL ecolume to determine the release of tritium label fromthe 5-position of the dUMP. A standard curve was established in theabsence of inhibitor, and three additional curves containing inhibitorat approximately 1/2 to 2 times the K_(i) were determined. Experimentalresults were analyzed by EZ-FIT, a nonlinear regression analysis program(Perrella Scientific, Springfield, Pa.) which was used to fit all datapoints simultaneously to a mixed noncompetitive inhibition scheme. Theresults obtained are shown in the Table. The first entry for eachcompound is the K_(i), slope and the entry underneath is the K_(i),intercept.

In Vitro Testing to Determine Inhibition of Growth of Tumor Cells

Cellular growth in the presence of the compounds in question wasassessed using three cell lines: the L1210 murine leukemia (ATCC CCL219), CCFR-CEM, a human lymphoblastic leukemia line of T-cell origin(ATCC CCL 119), and a thymidine kinase-deficient human colonadenocarcinoma, GC₃ /M TK⁻ (supplied by Drs. P. J. and J. A. Houghton,St. Jude Childrens Research Hospital, Memphis, Tenn.). Cell lines weremaintained in RPMI 1640 medium containing 5% (L1210, CCRF-CEM) or 10%(GC₃ /M TK⁻) heat-inactivated fetal bovine serum without antibiotics.

IC₅₀ values were determined in 150 μL microcultures each containing 1500(L1210) or 10,000 (CCRF-CEM, GC₃ /M TK⁻) cells established in 96 wellplates in growth medium supplemented with 50 U/mL penicillin and 50μg/mL streptomycin. Growth was measured over 3 days (L1210) or 5 days(CCRF-CEM, GC₃ /M TK⁻) of continuous exposure to varying concentrationsof each test compound, added 4 h. after initial cell plating, by theMTT-tetrazolium reduction assay of T. J. Mosmann [J. Immunol. Meth. 6555 (1983)] modified according to Alley et al. [Cancer Res. 48 589(1988)]. Water insoluble derivatives were dissolved in DMSO and dilutedto a final concentration of 0.5% solvent in cell cultures.

The results obtained from this procedure are-shown below in the Table 2.[Although Table 2 indicates that certain compounds do not demonstrateparticularly good TS inhibition, these compounds are of potentialinterest in that they may demonstrate other antitumor, activity such astoxicity to L1210 cells in tissue culture.]

                                      TABLE 2                                     __________________________________________________________________________    KiData (μM)    Cell Culture (IC50 μM)                                   E. coli  Human    L1210   CCRF-CEM                                                                              GC3-M (TK-)                                 __________________________________________________________________________     1A                                                                              >100  >100     --      --      --                                           2A                                                                              >10   >10      --      --      --                                           3A                                                                               >3    >3      --      10% > 12.3                                                                            20% > 12.7                                   4A                                                                              --    --       --      --      --                                           5A                                                                              --    --       --      --      --                                           6A                                                                              >10   >10      2.3     4.0     >4.98                                        7A                                                                              38 ± 5                                                                           2.1 ± 0.5                                                                           14      37% @ > 26.9                                                                          none @ > 26.9                                                 68% @ 20 um                                                  8A                                                                              0.89 ± 0.28                                                                      0.062 ± 0.23                                                                        3.5     5.2     6.0                                          9A                                                                              0.22 ± 0.07                                                                      0.13 ± 0.04                                                                         1.8     2.1     4.5                                         10A                                                                              0.75 ± 0.08                                                                      0.083 ± 0.011                                                                       3.0     2.9     >4.0                                        11A                                                                              21 ± 13                                                                          2.0 ± 0.4                                                                           none @ > 3.33                                                                         17% @ > 3.33                                                                          none @ > 3.33                               12A                                                                              0.55 ± 0.05                                                                       0.07 ± 0.001                                                                       4.2     4.2     5.0                                         13A                                                                              3.9 ± 0.9                                                                        0.64 ± 0.01                                                                         21      26      32                                          14A                                                                              0.15 ± 0.03                                                                      0.017 ± 0.008                                                                       1.0     0.81    1.0                                         15A                                                                              190 ± 130                                                                        19 ± 11                                                                             27% @ > 50                                                                            40% @ > 50                                                                             4% @ > 50                                  16A                                                                              0.76 ± 0.12                                                                      0.048 ± 0.006                                                                       3.1     3.8     >5.0                                        17A                                                                              0.54 ± 0.07                                                                      0.13 ± 0.03                                                                         8.1     8.6     15.0                                        18A                                                                              --    --       1.8     33% @ > 2.0                                                                           33% @ > 2.0                                 19A                                                                              --    --       --      --      --                                          20A                                                                              311 ±  99                                                                        61 ± 17                                                                             40% @ > 50                                                                            20% @ > 50                                                                            none @ > 50                                 21A                                                                              >80   190 ± 25                                                                            28% @ > 50                                                                            42% @ > 50                                                                            none @ > 50                                 22A                                                                              9.3 ± 1.6                                                                        1.1 ± 0.3                                                                             4% @ > 2.5                                                                          none @ > 2.5                                                                          none @ > 2.5                                23A                                                                              --     0.13 ± 0.009                                                                       3.5     5.1     6.1                                         24A                                                                              --    0.023 ± 0.001                                                                       0.55    1.1     1.2                                         25A                                                                              --    0.022 ± 0                                                                           0.59    1.1     1.7                                         26A                                                                              --    0.079 ± 0                                                                           4.05    10.5    18.0                                        27A                                                                              --    0.00795 ± 0                                                                         1.05    0.99    4.1                                         28A                                                                              --    0.115 ± 0                                                                           27% @ > 50                                                                            none @ > 50                                                                           none @ > 50                                 29A                                                                              1.1 ± 0.2                                                                        0.12 ± 0.02                                                                         8.0     10.5    >12.5                                       30A                                                                              0.14 ± 0                                                                         0.011 ± 0                                                                           1.6     0.88    1.5                                         31A                                                                              21.0 ± 6                                                                         51.0 ± 2.2                                                                          48.0    >50     > 50                                        32A                                                                              >10   >10      >10     6.0     30% @ > 10                                  33A                                                                               36 ± 1.5                                                                        47 ± 17                                                                             25.0    18.0    20% @ > 25                                  __________________________________________________________________________     * -- (Assay not performed)                                               

While the invention has been described in detail and with reference tospecific embodiments thereof, it will be apparent to one skilled in theart that various changes and modifications can be made therein withoutdeparting from the spirit and scope thereof. Thus, it is intended thatthe present invention cover the modifications and variations, providedthey come within the scope of the appended claims and their equivalents.

We claim:
 1. A process for making a quinazoline compound having theformula I ##STR74## wherein: R¹ represents hydrogen, halogen, alkyl,--OH, --O-alkyl, --O-(aryl or heteroaryl), --S-alkyl, --S-(aryl orheteroaryl), --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO, --NHOH,--NHO-alkyl, --NHNH₂, substituted --NHNH₂, --NHC(═NH)NH₂,--NHC(═NH)alkyl, fluoroalkyl, cycloalkyl, alkenyl, alkynyl, aryl, orheterocycle;R² and R³, which may be the same or different, representhydrogen, halogen, alkyl, cycloalkyl, --OH, --O-alkyl, --S-alkyl, --NH₂,--NH-alkyl, --N-(alkyl)₂, --NHCHO, --NO₂, --NHOH, --NHO-alkyl, --NHNH₂,substituted --NHNH₂, --CN, --CO₂ H, --CO₂ -alkyl, --CONH₂, --CONH-alkyl,--CON(alkyl)₂, --CSNH₂, --CSNH-alkyl, --CSN(alkyl)₂, --C(═NH)NH₂,--NHC(═NH)NH₂, --NHC(═NH)alkyl, --SO-alkyl, --SO₂ -alkyl, fluoroalkyl,--O-fluoroalkyl, --S-fluoroalkyl, --NHCO(alkyl), --NHCO(fluoroalkyl),--SO-fluoroalkyl, --SO₂ -fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂NH(alkyl), --SO₂ N(alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle; Zrepresents O or S; R⁴ represents O, S, SO, SO₂, NH, N-alkyl, CH₂,CH-alkyl, CH-(aryl or heteroaryl), CHOH, CHO-alkyl, CHO-(aryl orheteroaryl), C(alkyl)₂, C(aryl or heteroaryl)₂, C(alkyl)(aryl orheteroaryl), CHS-alkyl, CHS-aryl, C(OH)(alkyl), C(OH)(aryl orheteroaryl), C(OH)(cycloalkyl), N(OH), N-cycloalkyl, N(aryl orheteroaryl), C(cycloalkyl)₂, C(aryl or heteroaryl)(cycloalkyl),C(alkyl)(alkenyl), C(alkyl)(alkynyl), C(alkenyl)₂, C(alkynyl)₂,C(alkynyl)(aryl or heteroaryl), C(alkynyl)(alkenyl), C(alkenyl)(aryl orheteroaryl), C(cycloalkyl)(alkenyl), C(cycloalkyl)(alkynyl),C(alkyl)(aryl or heteroaryl), CH(cycloalkyl), CH(alkenyl), CH(alkynyl),C(alkyl)(cycloalkyl), C(alkyl)(O-alkyl), C(alkenyl)(O-alkyl),C(alkynyl)(O-alkyl), C(alkyl)(O-cycloalkyl), C(alkenyl)(O-cycloalkyl),C(alkynyl)(O-cycloalkyl), C(aryl or heteroaryl)(O-alkyl), C(aryl orheteroaryl)(O-cycloalkyl), C(alkynyl)(S-alkyl),C(alkynyl)(S-cycloalkyl), C(alkenyl)(S-alkyl), C(alkenyl)(S-cycloalkyl),C(alkyl)(S-alkyl), C(alkyl)(S-cycloalkyl), C(aryl orheteroaryl)(S-alkyl), C(aryl or heteroaryl)(S-cycloalkyl), N(NH₂),N[NH(alkyl)], N[N(alkyl)₂ ], N[NH(cycloalkyl)], N[N(alkyl)(cycloalkyl)],CH(NH₂), CH[NH(alkyl)], CH[NH(cycloalkyl)], CH[N(alkyl)₂ ],CH[N(alkyl)(cycloalkyl)], CH[N(cycloalkyl)₂ ], C(alkyl)(NH₂),C(alkyl)[NH(alkyl)], C(alkyl)[N(cycloalkyl)₂ ],C(alkyl)[NH(cycloalkyl)], C(alkyl)[N(alkyl)₂ ],C(alkyl)[N(alkyl)(cycloalkyl)], C(aryl or heteroaryl)(NH₂), C(aryl orheteroaryl)[NH(alkyl)], C(aryl or heteroaryl)[NH(cycloalkyl)], C(aryl orheteroaryl)[N(alkyl)₂ ], C(aryl or heteroaryl)[N(cycloalkyl)₂ ], orC(aryl or heteroaryl)[N(alkyl)(cycloalkyl)]; and R⁵ represents asubstituted or unsubstituted aryl or heteroaryl group which processcomprises subjecting a compound having the formula ##STR75## wherein Lis a leaving group, to a displacement reaction with the appropriatecompound which will cause the leaving group L to be replaced with thedesired --R4--R⁵ substituent in formula (i) or with the appropriate --R⁵substituent in formula (ii).
 2. A process for making a compound of theformula ##STR76## wherein R² and R³, which may be the same or different,represent hydrogen, halogen, alkyl, cycloalkyl, --OH, --O-alkyl,--S-alkyl, --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO, --NO₂, --NHOH,--NHO-alkyl, --NHNH₂, substituted --NHNH₂, --CN, --CO₂ H, --CO₂ -alkyl,--CONH₂, --CONH-alkyl, --CON(alkyl)₂, --CSNH₂, --CSNH-alkyl,--CSN(alkyl)₂, --C(═NH)NH₂, --NHC(═NH) NH₂, --NHC(═NH)alkyl, --SO-alkyl,--SO₂ -alkyl, fluoroalkyl, --O-fluoroalkyl, --S-fluoroalkyl,--NHCO(alkyl), --NHCO(fluoroalkyl), --SO-fluoroalkyl, --SO₂-fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂ NH(alkyl), --SO₂N(alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle;R⁴ represents O, S,SO, SO₂, NH, N-alkyl, CH₂, CH-alkyl, CH-(aryl or heteroaryl), CHOH,CHO-alkyl, CHO-(aryl or heteroaryl), C(alkyl)₂, C(aryl or heteroaryl)₂,C(alkyl)(aryl or heteroaryl), CHS-alkyl, CHS-aryl, C(OH)(alkyl),C(OH)(aryl or heteroaryl), C(OH)(cycloalkyl), N(OH), N-cycloalkyl,N(aryl or heteroaryl), C(cycloalkyl)₂, C(aryl orheteroaryl)(cycloalkyl), C(alkyl)(alkenyl), C(alkyl)(alkynyl),C(alkenyl)₂, C(alkynyl)₂, C(alkynyl)(aryl or heteroaryl),C(alkynyl)(alkenyl), C(alkenyl)(aryl or heteroaryl),C(cycloalkyl)(alkenyl), C(cycloalkyl)(alkynyl), C(alkyl)(aryl orheteroaryl), CH(cycloalkyl), CH(alkenyl), CH(alkynyl),C(alkyl)(cycloalkyl), C(alkyl)(O-alkyl), C(alkenyl)(O-alkyl),C(alkynyl)(O-alkyl), C(alkyl)(O-cycloalkyl), C(alkenyl)(O-cycloalkyl),C(alkynyl)(O-cycloalkyl), C(aryl or heteroaryl)(O-alkyl), C(aryl orheteroaryl)(O-cycloalkyl), C(alkynyl)(S-alkyl),C(alkynyl)(S-cycloalkyl), C(alkenyl)(S-alkyl), C(alkenyl)(S-cycloalkyl),C(alkyl)(S-alkyl), C(alkyl)(S-cycloalkyl), C(aryl orheteroaryl)(S-alkyl), C(aryl or heteroaryl)(S-cycloalkyl), N(NH₂),N[NH(alkyl)], N[N(alkyl)₂ ], N[NH(cycloalkyl)], N[N(alkyl)(cycloalkyl)],CH(NH₂), CH[NH(alkyl)], CH[NH(cycloalkyl)], CH[N(alkyl)₂ ],CH[N(alkyl)(cycloalkyl)], CH[N(cycloalkyl)₂ ], C(alkyl)(NH₂),C(alkyl)[NH(alkyl)], C(alkyl)[N(cycloalkyl)₂ ],C(alkyl)[NH(cycloalkyl)], C(alkyl)[N(alkyl)₂ ],C(alkyl)[N(alkyl)(cycloalkyl)], C(aryl or heteroaryl)(NH₂), C(aryl orheteroaryl)[NH (alkyl)], C(aryl or heteroaryl)[NH(cycloalkyl)], C(arylor heteroaryl)[N(alkyl)₂ ], C(aryl or heteroaryl)[N(cycloalkyl)₂ ], orC(aryl or heteroaryl)[N(alkyl)(cycloalkyl)]; and R⁵ represents asubstituted or unsubstituted aryl or heteroaryl group which processcomprises the steps of:(1) reacting a compound of the formula ##STR77##wherein L is a leaving group, with a hydroxylamine hydrochloride to forman isonitrosoacetanilide compound of the formula ##STR78## (2) treatingthe isonitrosoacetanilide compound of step (1) with acid, followed bythe addition of ice and purification with alcohol, to obtain an isatincompound of the formula ##STR79## (3) reacting the isatin compound ofstep (2) with an aqueous basic peroxide to form an anthranilic acidcompound of the formula ##STR80## (4) reacting the anthranilic acidcompound of step (3) with acetic anhydride to form an acetylanthranilcompound of the formula ##STR81## (5) reacting the acetylanthranilcompound of step (4) with ammonia, followed by treatment with a base andthen by acid to obtain a quinazoline of the formula ##STR82## (6)subjecting the quinazoline of step (5) to a displacement reaction toreplace the leaving group L with the desired --R⁴ --R⁵ substituent toobtain said compound of the formula ##STR83##
 3. A process for making acompound of the formula ##STR84## wherein R² and R³, which may be thesame or different, represent hydrogen, halogen, alkyl, cycloalkyl, --OH,--O-alkyl, --S-alkyl, --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO, --NO₂,--NHOH, --NHO-alkyl, --NHNH₂, substituted --NHNH₂, --CN, --CO₂ H, --CO₂-alkyl, --CONH₂, --CONH-alkyl, --CON(alkyl)₂, --CSNH₂, --CSNH-alkyl,--CSN(alkyl)₂, --C(═NH)NH₂, --NHC(═NH)NH₂, --NHC(═NH)alkyl, --SO-alkyl,-SO₂ -alkyl, fluoroalkyl, --O-fluoroalkyl, --S-fluoroalkyl,--NHCO(alkyl), --NHCO(fluoroalkyl), --SO-fluoroalkyl, --SO₂-fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂ NH(alkyl), --SO₂N(alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle;and L is a leavinggroup; which process comprises the steps of:(1) reacting a compound ofthe formula ##STR85## wherein L is a leaving group, with a hydroxylaminehydrochloride to form an isonitrosoacetanilide compound of the formula##STR86## (2) treating the isonitrosoacetanilide compound of step (1)with acid, followed by addition of ice and purification with alcohol, toobtain an isatin compound of the formula ##STR87## (3) reacting theisatin compound of step (2) with an aqueous basic peroxide to form ananthranilic acid compound of the formula ##STR88## (4) reacting theanthranilic acid compound of step (3) with acetic anhydride to form anacetylanthranil compound of the formula ##STR89## (5) treating theacetylanthranil compound of step (4) with alcohol, followed by acid, toobtain a compound of the formula ##STR90## and (6) reacting the productof step (5) with chloroformamidine hydrochloride to obtain a quinazolinecompound of the formula ##STR91##
 4. A process of making a compound ofthe formula ##STR92## wherein R² and R³, which may be the same ordifferent, represent hydrogen, halogen, alkyl, cycloalkyl, --OH,--O-alkyl, --S-alkyl, --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO, --NO₂,--NHOH, --NHO-alkyl, --NHNH₂, substituted --NHNH₂, --CN, --CO₂ H, --CO₂-alkyl, --CONH₂, --CONH-alkyl, --CON(alkyl)₂, --CSNH₂, --CSNH-alkyl,--CSN(alkyl)₂, --C(═NH)NH₂, --NHC(═NH)NH₂, --NHC(═NH)alkyl, --SO-alkyl,--SO₂ -alkyl fluoroalkyl, --O-fluoroalkyl, --S-fluoroalkyl,--NHCO(alkyl), --NHCO(fluoroalkyl), --SO-fluoroalkyl, --SO₂-fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂ NH(alkyl), --SO₂N(alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle;and L is a leavinggroup; which process comprises the steps of:(1) reacting a compound ofthe formula ##STR93## wherein L is a leaving group, with a hydroxylaminehydrochloride to form an isonitrosoacetanilide compound of the formula##STR94## (2) treating the isonitrosoacetanilide compound of step (1)with acid, followed by addition of ice and purification with alcohol, toobtain an isatin compound of the formula ##STR95## (3) reacting theisatin compound of step (2) with an aqueous basic peroxide to form ananthranilic acid compound of the formula ##STR96## (4) reacting theanthranilic acid compound of step (3) with phosgene or triphosgene toform a compound of the formula ##STR97## which is further reacted withalcohol; (5) reacting the product of step (4) with chloroformamidinehydrochloride to obtain a quinazoline compound of the formula ##STR98##5. A process for preparing a compound of the formula ##STR99## whereinR¹ represents hydrogen, halogen, alkyl, --OH, --O-alkyl, --O-(aryl orheteroaryl), --S-alkyl, --S-(aryl or heteroaryl), --NH₂, --NH-alkyl,--N-(alkyl)₂, --NHCHO, --NHOH, --NHO-alkyl, --NHNH₂, substituted--NHNH₂, --NHC(═NH)NH₂, --NHC(═NH) alkyl, fluoroalkyl, cycloalkyl,alkenyl, alkynyl, aryl, or heterocycle;R² and R³, which may be the sameor different, represent hydrogen, halogen, alkyl, cycloalkyl, --OH,--O-alkyl, --S-alkyl, --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO, --NO₂,--NHOH, --NHO-alkyl, --NHNH₂, substituted --NHNH₂, --CN, --CO₂ H, --CO₂-alkyl, --CONH₂, --CONH-alkyl, --CON(alkyl)₂, --CSNH₂, --CSNH-alkyl,--CSN(alkyl)₂, --C(═NH)NH₂, --NHC(═NH)NH₂, --NHC(═NH)alkyl, --SO-alkyl,--SO₂ -alkyl, fluoroalkyl, --O-fluoroalkyl, --S-fluoroalkyl,--NHCO(alkyl), --NHCO(fluoroalkyl), --SO-fluoroalkyl, --SO₂-fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂ NH(alkyl), --SO₂N(alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle; Z represents O or S;and R⁹ represents hydrogen, halogen, --CH₃, --OCH₃, --CF₃, or N(CH₃),which process comprises the steps of:(1) reacting a compound of theformula ##STR100## with a benzyl mercaptan to form a compound of theformula ##STR101## wherein R¹⁰ is hydrogen or --OCH₃ ; (2) reducing theproduct of step (1); (3) deprotecting the product of step (2); and (4)reacting the product of step (3) with a compound of the formula##STR102## wherein L is a leaving group, to obtain a compound of theformula ##STR103##
 6. A process for preparing a compound of the formula##STR104## wherein R¹ represents hydrogen, halogen, alkyl, --OH,--O-alkyl, --O-(aryl or heteroaryl), --S-alkyl, --S-(aryl orheteroaryl), --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO, --NHOH,--NHO-alkyl, --NHNH₂, substituted --NHNH₂, --NHC(═NH)NH₂,--NHC(═NH)alkyl, fluoroalkyl, cycloalkyl, alkenyl, alkynyl, aryl, orheterocycle;R² and R³, which may be the same or different, representhydrogen, halogen, alkyl, cycloalkyl, --OH, --O-alkyl, --S-alkyl, --NH₂,--NH-alkyl, --N-(alkyl)₂, --NHCHO, --NO₂, --NHOH, --NHO-alkyl, --NHNH₂,substituted --NHNH₂, --CN, --CO₂ H, --CO₂ -alkyl, --CONH₂, --CONH-alkyl,--CON(alkyl)₂, --CSNH₂, --CSNH-alkyl, --CSN(alkyl)₂, --C(═NH)NH₂,--NHC(═NH)NH₂, --NHC(═NH)alkyl, --SO-alkyl, --SO₂ -alkyl, fluoroalkyl,--O-fluoroalkyl, --S-fluoroalkyl, --NHCO(alkyl), --NHCO(fluoroalkyl),--SO-fluoroalkyl, --SO₂ -fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂NH(alkyl), --SO₂ N(alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle; Zrepresents O or S; and R⁹ represents hydrogen, halogen, --CH₃, --OCH₃,--CF₃, or N(CH₃), which process comprises the steps of:(1) reducing acompound of the formula ##STR105## to form a compound of the formula##STR106## (2) reacting the product of step (1) with a xanthate compoundto obtain a compound of the formula ##STR107## and (3) subjecting theproduct of step (2) to hydrolysis and reaction with a compound of theformula ##STR108## wherein L is a leaving group, in the presence ofN,N-dimethylacetamide, copper (I) bromide and copper (I) oxide to obtaina compound of the formula ##STR109##
 7. A process for preparing acompound of the formula ##STR110## wherein R¹ represents hydrogen,halogen, alkyl, --OH, --O-alkyl, --O-(aryl or heteroaryl), --S-alkyl,--S-(aryl or heteroaryl), --NH₂, --NH-alkyl, --N-(alkyl)₂, --NHCHO,--NHOH, --NHO-alkyl, --NHNH₂, substituted --NHNH₂, --NHC(═NH)NH₂,--NHC(═NH)alkyl, fluoroalkyl, cycloalkyl, alkenyl, alkynyl, aryl, orheterocycle;R² and R³, which may be the same or different, representhydrogen, halogen, alkyl, cycloalkyl, --OH, --O-alkyl, --S-alkyl, --NH₂,--NH-alkyl, --N-(alkyl)₂, --NHCHO, --NO₂, --NHOH, --NHO-alkyl, --NHNH₂,substituted --NHNH₂, --CN, --CO₂ H, --CO₂ -alkyl, --CONH₂, --CONH-alkyl,--CON(alkyl)₂, --CSNH₂, --CSNH-alkyl, --CSN(alkyl)₂, --C(═NH)NH₂,--NHC(═NH)NH₂, --NHC(═NH)alkyl, --SO-alkyl, --SO₂ -alkyl, fluoroalkyl,--O-fluoroalkyl, --S-fluoroalkyl, --NHCO(alkyl), --NHCO(fluoroalkyl),--SO-fluoroalkyl, --SO₂ -fluoroalkyl, --SH, --SO₃ H, --SO₂ NH₂, --SO₂ NH(alkyl), --SO₂ N(alkyl)₂, alkenyl, alkynyl, aryl, or heterocycle; Zrepresents O or S; R¹⁰ is hydrogen or --OH; R¹¹ is hydrogen or aryl; andR⁵ represents a substituted or unsubstituted aryl or heteroaryl groupwhich process comprises the steps of(1) reacting a compound of theformula ##STR111## with a compound suitable for providing a protectinggroup P to form a compound of the formula ##STR112## (2) converting theproduct of step (1) to a compound of the formula ##STR113## wherein L isa leaving group; (3) converting the product of step (2) to an aldehydeof the formula ##STR114## (4) subjecting the aldehyde compound of step(3) to a reaction to form a compound of the formula ##STR115## whereinR¹⁰ is --OH and R¹¹ is hydrogen; and (5) deprotecting the product ofstep (4) to obtain a compound of the formula ##STR116## wherein R¹⁰ is--OH and R¹¹ is hydrogen.
 8. A process according to claim 1 wherein acompound having the formula (ii) is subjected to said displacementreaction.
 9. A process of making a compound according to claim 1,wherein said process is carried out in the presence of a base at atemperature of from about 70° C. to about 165° C.
 10. A process ofmaking a compound according to claim 2, wherein step (1) is carried inthe presence of water, chloral hydrate, HCl, sodium sulfate andhydroxylamine hydrochloride at a temperature of from about 0° C. toabout 100° C.
 11. A process of making a compound according to claim 2,wherein step (2) is carried in the presence of concentrated H₂ SO₄ at atemperature of from about 50° C. to about 100° C.
 12. A process ofmaking a compound according to claim 2, wherein step (3) is carried inthe presence of basic, aqueous hydrogen peroxide at a temperature offrom about 0° C. to about 80° C.
 13. A process of making a compoundaccording to claim 2, wherein step (4) is carried in the presence ofacetic anhydride at a temperature of from about 70° C. to about 140° C.14. A process of making a compound according to claim 2, wherein step(5) is carried in the presence of ammonia at a temperature of from about-33° C. to about 20° C.
 15. A process of making a compound according toclaim 2, wherein step (6) is carried out in the presence of a base andappropriate catalyst at a temperature of from about 70° C. to about 165°C.
 16. A process of making a compound according to claim 5, wherein step(2) is carried out in the presence of a solvent and a reducing agent ata temperature of from about 0° C. to about 80° C.
 17. A process ofmaking a compound according to claim 5, wherein step (3) is carried outin the presence of a solvent and a metal or metal salt at a temperatureof from about -78° C. to about 20° C.
 18. A process of making a compoundaccording to claim 5, wherein step (4) is carried out in the presence ofsodium hydride, N,N-dimethylacetamide, copper (I) bromide and copper (i)oxide at a temperature of from about 70° C. to about 165° C.
 19. Aprocess of making a compound according to claim 6, wherein step (1) iscarried out in the presence of hydrogen gas, a solvent and a catalyticamount of palladium, at a temperature of about 20° C.
 20. A process ofmaking a compound according to claim 6, wherein step (2) is carried outin the presence of an aqueous acid, followed by NaNO₂ and potassiumxanthate, at a temperature of from about -40° C. to about 20° C.
 21. Aprocess of making a compound according to claim 6, wherein thehydrolysis of step (3) is carried out in the presence NaOH/CH₃ OH at atemperature of from about 0° C. to about 20° C.
 22. A process of makinga compound according to claim 6, wherein the reaction part of step (3)is carried out in the presence of a base, solvent and catalyst at atemperature of from about 70° C. to about 165° C.
 23. A process ofmaking a compound according to claim 7, wherein step (1) is carried outin the presence of an alkyl or acyl halide, a base and a solvent at atemperature of from about 0° C. to about 20° C.
 24. A process of makinga compound according to claim 7, wherein said protecting group P isselected from the group consisting of CH₂ OCH₂ CH₂ Si(CH₃)₃, CH₂ OCH₃,CH₂ OC(O)^(t) Bu and CO^(t) Bu.
 25. A process of making a compoundaccording to claim 24, wherein said protecting group P is CH₂ OCH₂ CH₂Si(CH₃)₃.
 26. A process of making a compound according to claim 7,wherein step (2) is carried out in the presence of N-Bromosuccinimide,bromine, N-Chlorosuccinimide or N-Iodosuccinimide.
 27. A process ofmaking a compound according to claim 26, wherein step (2) is carried outin the presence of N-Bromosuccinimide.
 28. A process of making acompound according to claim 27, wherein step (2) is carried out in thepresence of N-Bromosuccinimide, CCl₄ and light at a temperature of fromabout 50° C. to about 100° C.
 29. A process of making a compoundaccording to claim 7, wherein step (3) is carried out in the presence ofa nucleophile, base and solvent at a temperature of from about 0° C. toabout 100° C.
 30. A process of making a compound according to claim 7,wherein step (3) is carried out by reacting the product of step (2) with5-chloroindole.
 31. A process of making a compound according to claim 7wherein step (3) is carried out by reacting the product of step (2) withNaOEt and 2-nitropropane and further wherein the product of step (3) isreacted with phenylmagnesium in step (4) to form a compound of theformula ##STR117## wherein R¹⁰ is --OH and R¹¹ is phenyl.
 32. A processof making a compound according to claim 7, wherein step (5) is carriedout in the presence of an acid or basic fluoride at a temperature offrom about 0° C. to about 100° C.
 33. A process of making a compoundaccording to claim 7, wherein deprotecting step (5) is carried out byreacting the product of step (5) with tetrabutylammonium fluoride.
 34. Aprocess of making a compound according to claim 7, wherein:(i) prior todeprotecting step (5), the product of step (4) is oxidized to form acompound of the formula ##STR118## wherein R⁵ is phenyl; and (ii)subsequent to deprotecting step (5), the oxidation product is reactedwith phenyllithium to form a compound of the formula ##STR119## whereinR⁵ is phenyl, R¹¹ is phenyl and R¹⁰ is --OH.
 35. A process of making acompound according to claim 3, wherein step (5) is carried out (i) inthe presence of methanol at a temperature of from about 0° C. to about100° C. and then (ii) in the presence of concentrated HCl at atemperature of from about 70° C. to about 100° C.
 36. A process ofmaking a compound according to claim 3, wherein step (4) is carried out(i) in the presence of triphosgene at a temperature of from about 0° C.to about 20° C. and then (ii) in the presence of methanol at atemperature of from about 0° C. to about 20° C.
 37. A process of makinga compound according to claim 3, wherein step (6) is carried out in thepresence of diglyme and cloroformamidine hydrochloride at a temperatureof from about 160° C. to about 175° C.
 38. A process of making acompound according to claim 2, wherein step (6) is carried out byreacting the product of step (5) with a 4-thiopyridine anion, in thepresence of sodium hydride, copper (I) bromide and copper (I) oxide. 39.A process of making a compound according to claim 6, wherein step (3) iscarried out by reacting the product of step (2) with a 4-thiopyridineanion, in the presence of sodium hydride, copper (I) bromide and copper(I) oxide.
 40. A process of making a compound according to claim 38,wherein the 4-thiopyridine anion is made by reacting 4-mercaptopyridinewith NaH in anhydrous N,N-Dimethylacetamide.
 41. A process of making acompound according to claim 39, wherein the 4-thiopyridine anion is madeby reacting 4-mercaptopyridine with anhydrous N,N-Dimethylacetamide inNaH.
 42. A process of making a compound according to claim 5, whereinstep (1) is carried out in the presence of a base and solvent at atemperature of from about 0° C. to about 80° C.